Crystalline C21H22Cl2N4O2 malonate

ABSTRACT

The present invention provides a malonate salt of a compound of formula (I): 
                         
which is a crystalline salt. Also provided are pharmaceutical compositions that include the provided malonate salt and methods of using the provided crystalline forms and pharmaceutical compositions for the treatment of cancer.

CROSS-REFERENCE TO RELATED APPLICATIONS

This patent application claims the benefit of U.S. Provisional PatentApplication No. 62/110,446, filed Jan. 30, 2015, which is incorporatedby reference herein in its entirety.

FIELD OF THE INVENTION

The present invention relates to crystalline4-(5-Chloro-2-isopropylaminopyrid-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate, which is useful as aninhibitor of ERK protein kinase.

BACKGROUND OF THE INVENTION

Mitogen-activated protein kinase (MAPK) pathways mediate signals whichcontrol diverse cellular processes including growth, differentiation,migration, proliferation and apoptosis. One MAPK pathway, theextracellular signal-regulated kinase (ERK) signaling pathway, is oftenfound to be up-regulated in tumors. Pathway members, therefore,represent attractive blockade targets in the development of cancertherapies (Kohno and Pouyssegur, 2006). For example, U.S. Pat. No.7,354,939 B2 discloses, inter alia, compounds effective as inhibitors ofERK protein kinase. One of these compounds,4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide, is a compound according toformula (I):

Pharmaceutical compositions are often formulated with a crystallinesolid of the active pharmaceutical ingredient (API). The specificcrystalline form of the API can have significant effects on propertiessuch as stability and solubility/bioavailability. Instability andsolubility characteristics can limit the ability to formulate acomposition with an adequate shelf life or to effectively deliver adesired amount of a drug over a given time frame. One strategy used toachieve the desired physical parameters is the practice of saltselection. (Peterson et al., 2006).

There exists an unmet need for crystalline forms of4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide which exhibit improvedproperties for formulation of pharmaceutical compositions. The presentapplication is directed to meeting this and other needs.

SUMMARY OF THE INVENTION

It has been discovered that crystalline forms of4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonalte can be prepared whichexhibit improved properties, e.g. surprisingly improved stability andimproved solubility characteristics.

Thus, the present invention provides a malonate salt of a compound offormula (I):

which is a crystalline salt.

The present invention also provides a malonate salt of a compound offormula (I):

having an XRPD pattern substantially as shown in FIG. 7.

The present invention also provides Form A crystalline4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate having an infraredspectroscopy (IR) spectrum comprising one or more peaks at about 1573,1504, 1475, 1253, 1033, and 883 cm⁻¹.

The present invention also provides Form A crystalline4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate having an IR spectrumsubstantially as shown in FIG. 8.

The present invention also provides Form A crystalline4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate having (i) an XRPDpattern comprising one or more peaks at about 3.0. 5.2, 8.0, and 10.9°2θ; and (ii) an IR spectrum comprising one or more peaks at about 1573,1504, 1475, 1253, 1033, and 883 cm⁻¹.

The present invention also provides Form A crystalline4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate having having a DSCthermogram with an endotherm having an onset temperature ofapproximately 142.1° C.

The present invention also provides Form A crystalline4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate having a DSCthermogram substantially as shown in FIG. 9.

The present invention also provides a pharmaceutical compositioncomprising a crystalline compound of the present invention and apharmaceutically acceptable carrier.

The present invention also provides a method of treating a cancer in asubject in need thereof comprising administering to the subject aneffective amount of a crystalline compound of the present invention.

The present invention also provides a method of treating a cancer in asubject in need thereof comprising administering to the subject aneffective amount of a pharmaceutical composition of the presentinvention.

The present invention also provides a method of making Form Acrystalline4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate comprising reactingmalonic acid and4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide under conditions suitable toform Form A crystalline4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate.

BRIEF DESCRIPTION OF THE DRAWINGS

The following drawings form part of the present specification and areincluded to further demonstrate certain aspects of the presentinvention. The invention may be better understood by reference to one ormore of these drawings in combination with the detailed description ofspecific embodiments presented herein.

FIG. 1 shows the XRPD of4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide free base acquired intransmission mode.

FIG. 2 shows the FT-IR spectrum of4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide free base.

FIG. 3 shows the DSC thermogram of4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide free base.

FIG. 4 shows the XRPD of4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide Form C acquired in transmissionmode.

FIG. 5 shows the FT-IR spectrum of4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide Form C.

FIG. 6 shows the DSC thermogram of4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide Form C.

FIG. 7 shows the XRPD of4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate Form A acquired intransmission mode.

FIG. 8 shows the FT-IR spectrum of4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate Form A.

FIG. 9 shows the DSC thermogram of4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate Form A.

FIG. 10 shows solubility of4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide Form C (“Form C”) and malonateform A. FIG. 10A: solubility in fasting state simulated gastric fluid(FaSSGF) pH 1.6. FIG. 10B: solubility in fasting state simulatedintestinal fluid (FaSSIF) pH 6.5.

FIG. 11 shows pharmacokinetics of4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide Form C (“Form C”) and malonateform A. FIG. 11A: Plasma drug concentration following a single dose of 5mg/kg. FIG. 11B: Area under the curve and coefficient of variation forin vivo pharmacokinetic studies.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides a malonate salt of a compound of formula

which is a crystalline salt.

In some embodiments, the malonate salt of the present invention ischaracterized by an X-ray powder diffraction (XRPD) pattern comprising acharacteristic peak at about 3.0° 2θ.

In some embodiments, the malonate salt of the present invention ischaracterized by an X-ray powder diffraction (XRPD) pattern comprisingcharacteristic peaks at about 3.0 and 5.2° 2θ.

In some embodiments, the malonate salt of the present invention ischaracterized by an X-ray powder diffraction (XRPD) pattern comprisingcharacteristic peaks selected from the group consisting of about 3.0,5.2, 8.0, and 10.9° 2θ.

In some embodiments, the malonate salt of the present invention ischaracterized by an X-ray powder diffraction (XRPD) pattern comprisingXRPD 2θ-reflections (°) at about 3.0. 5.2, 8.0, 10.9, 15.7, 18.4, 23.1,and 25.4.

The present invention also provides a malonate salt of a compound offormula (I):

having an XRPD pattern substantially as shown in FIG. 7.

The present invention also provides Form A crystalline4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate having an infraredspectroscopy (IR) spectrum comprising one or more peaks at about 1573,1504, 1475, 1253, 1033, and 883 cm⁻¹.

The present invention also provides Form A crystalline4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate having an IR spectrumsubstantially as shown in FIG. 8.

The present invention also provides Form A crystalline4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate having (i) an XRPDpattern comprising one or more peaks at about 3.0. 5.2, 8.0, and 10.9°2θ; and (ii) an IR spectrum comprising one or more peaks at about 1573,1504, 1475, 1253, 1033, and 883 cm⁻¹.

The present invention also provides Form A crystalline4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate having a DSCthermogram with an endotherm having an onset temperature ofapproximately 142.1° C.

The present invention also provides Form A crystalline4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate having a DSCthermogram substantially as shown in FIG. 9.

The present invention also provides a pharmaceutical compositioncomprising a crystalline compound of the present invention and apharmaceutically acceptable carrier.

The present invention also provides a method of treating a cancer in asubject in need thereof comprising administering to the subject aneffective amount of a crystalline compound of the present invention.

In some embodiments, the subject is a mammal.

In some embodiments, the mammal is selected from the group consisting ofhumans, primates, farm animals, and domestic animals.

In some embodiments, the mammal is a human.

In some embodiments, the method further comprises administering to thesubject at least one additional anti-cancer agent.

The present invention also provides a method of treating a cancer in asubject in need thereof comprising administering to the subject aneffective amount of a pharmaceutical composition of the presentinvention.

In some embodiments, the subject is a mammal.

In some embodiments, the mammal is selected from the group consisting ofhumans, primates, farm animals, and domestic animals.

In some embodiments, the mammal is a human.

In some embodiments, the method further comprises administering to thesubject at least one additional anti-cancer agent.

The present invention also provides a method of making Form Acrystalline4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate comprising reactingmalonic acid and4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide under conditions suitable toform Form A crystalline4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate.

In some embodiments, the malonic acid and4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide are reacted in an ethanolslurry.

The term “solid form” is often used to refer to a class or type ofsolid-state material. One kind of solid form is a “polymorph” whichrefers to two or more compounds having the same chemical formula butdiffering in solid-state structure. Salts may be polymorphic. Whenpolymorphs are elements, they are termed allotropes. Carbon possessesthe well-known allotropes of graphite, diamond, andbuckminsterfullerene. Polymorphs of molecular compounds, such as activepharmaceutical ingredients (“APIs”), are often prepared and studied inorder to identify compounds meeting scientific or commercial needsincluding, but not limited to, improved solubility, dissolution rate,hygroscopicity, and stability.

Other solid forms include solvates and hydrates of compounds includingsalts. A solvate is a compound wherein a solvent molecule is present inthe crystal structure together with another compound, such as an API.When the solvent is water, the solvent is termed a hydrate. Solvates andhydrates may be stoichiometric or non-stoichiometric. A monohydrate isthe term used when there is one water molecule, stoichiometrically, withrespect to, for example, an API, in the unit cell.

In order to identify the presence of a particular solid form, one ofordinary skill typically uses a suitable analytical technique to collectdata on the form for analysis. For example, chemical identity of solidforms can often be determined with solution-state techniques such as¹³C-NMR or ¹H-NMR spectroscopy and such techniques may also be valuablein determining the stoichiometry and presence of “guests” such as wateror solvent in a hydrate or solvate, respectively. These spectroscopictechniques may also be used to distinguish, for example, solid formswithout water or solvent in the unit cell (often referred to as“anhydrates”), from hydrates or solvates.

Solution-state analytical techniques do not provide information aboutthe solid state as a substance and thus, for example, solid-statetechniques may be used to distinguish among solid forms such asanhydrates. Examples of solid-state techniques which may be used toanalyze and characterize solid forms, including anhydrates and hydrates,include single crystal X-ray diffraction, X-ray powder diffraction(“XRPD”), solid-state ¹³C-NMR, Infrared (“IR”) spectroscopy, includingFourier Transform Infrared (FT-IR) spectroscopy, Raman spectroscopy, andthermal techniques such as Differential Scanning calorimetry (DSC),melting point, and hot stage microscopy.

Polymorphs are a subset of crystalline forms that share the samechemical structure but differ in how the molecules are packed in asolid. When attempting to distinguish polymorphs based on analyticaldata, one looks for data which characterize the form. For example, whenthere are two polymorphs of a compound (e.g., Form I and Form II), onecan use X-ray powder diffraction peaks to characterize the forms whenone finds a peak in a Form I pattern at angles where no such peak ispresent in the Form II pattern. In such a case, that single peak forForm I distinguishes it from Form II and may further act to characterizeForm I. When more forms are present, then the same analysis is also donefor the other polymorphs. Thus, to characterize Form I against the otherpolymorphs, one would look for peaks in Form I at angles where suchpeaks are not present in the X-ray powder diffraction patterns of theother polymorphs. The collection of peaks, or indeed a single peak,which distinguishes Form I from the other known polymorphs is acollection of peaks which may be used to characterize Form I. If, forexample, two peaks characterize a polymorph then those two peaks can beused to identify the presence of that polymorph and hence characterizethe polymorph. Those of ordinary skill in the art will recognize thatthere are often multiple ways, including multiple ways using the sameanalytical technique, to characterize polymorphic polymorphs. Forexample, one may find that three X-ray powder diffraction peakscharacterize a polymorph. Additional peaks could also be used, but arenot necessary, to characterize the polymorph up to and including anentire diffraction pattern. Although all the peaks within an entirediffractogram may be used to characterize a crystalline form, one mayinstead, and typically does as disclosed herein, use a subset of thatdata to characterize such a crystalline form depending on thecircumstances.

For example, as used herein, “characteristic peaks” are a subset ofobserved peaks and are used to differentiate one crystalline polymorphfrom another crystalline polymorph. Characteristic peaks are determinedby evaluating which observed peaks, if any, are present in onecrystalline polymorph of a compound against all other known crystallinepolymorphs of that compound to within ±0.2° 2θ.

When analyzing data to distinguish an anhydrate from a hydrate, forexample, one can rely on the fact that the two solid forms havedifferent chemical structures—one having water in the unit cell and theother not. Thus, this feature alone may be used to distinguish the formsof the compound and it may not be necessary to identify peaks in theanhydrate, for example, which are not present in the hydrate or viceversa.

X-ray powder diffraction patterns are some of the most commonly usedsolid-state analytical techniques used to characterize solid forms. AnX-ray powder diffraction pattern is an x-y graph with the diffractionangle, 2θ (°), on the x-axis and intensity on the y-axis. The peakswithin this plot may be used to characterize a crystalline solid form.The data is often represented by the position of the peaks on the x-axisrather than the intensity of peaks on the y-axis because peak intensitycan be particularly sensitive to sample orientation (see PharmaceuticalAnalysis, Lee & Web, pp. 255-257 (2003)). Thus, intensity is nottypically used by those skilled in the art to characterize solid forms.

As with any data measurement, there is variability in X-ray powderdiffraction data. In addition to the variability in peak intensity,there is also variability in the position of peaks on the x-axis. Thisvariability can, however, typically be accounted for when reporting thepositions of peaks for purposes of characterization. Such variability inthe position of peaks along the x-axis derives from several sources. Onecomes from sample preparation. Samples of the same crystalline material,prepared under different conditions may yield slightly differentdiffractograms. Factors such as particle size, moisture content, solventcontent, and orientation may all affect how a sample diffracts X-rays.Another source of variability comes from instrument parameters.Different X-ray instruments operate using different parameters and thesemay lead to slightly different diffraction patterns from the samecrystalline solid form. Likewise, different software packages processX-ray data differently and this also leads to variability. These andother sources of variability are known to those of ordinary skill in thepharmaceutical arts.

Due to such sources of variability, it is common to recite X-raydiffraction peaks using the word “about” prior to the peak value in ° 2θ(sometimes expressed herein as “2θ-reflections (°)”), which presents thedata to within 0.1 or 0.2° 2θ of the stated peak value depending on thecircumstances. The X-ray powder diffraction data corresponding to thesolid forms of the present invention were collected on instruments whichwere routinely calibrated and operated by skilled scientists. In thepresent invention, XRPD values are preferably obtained using Cu Kα X-rayradiation according to the method described in Example 1. Accordingly,the variability associated with these data would be expected to becloser to ±0.1° 2θ than to ±0.2° 2θ and indeed likely less than 0.1 withthe instruments used herein. However, to take into account thatinstruments used elsewhere by those of ordinary skill in the art may notbe so maintained, for example, all X-ray powder diffraction peaks citedherein have been reported with a variability on the order of ±0.2° 2θand are intended to be reported with such a variability wheneverdisclosed herein and are reported in the specification to onesignificant figure after the decimal even though analytical output maysuggest higher precision on its face.

Single-crystal X-ray diffraction provides three-dimensional structuralinformation about the positions of atoms and bonds in a crystal. It isnot always possible or feasible, however, to obtain such a structurefrom a crystal, due to, for example, insufficient crystal size ordifficulty in preparing crystals of sufficient quality forsingle-crystal X-ray diffraction.

X-ray powder diffraction data may also be used, in some circumstances,to determine the crystallographic unit cell of the crystallinestructure. The method by which this is done is called “indexing.”Indexing is the process of determining the size and shape of thecrystallographic unit cell consistent with the peak positions in asuitable X-ray powder diffraction pattern. Indexing provides solutionsfor the three unit cell lengths (a, b, c), three unit cell angles (α, β,γ), and three Miller index labels (h, k, l) for each peak. The lengthsare typically reported in Angstrom units and the angles in degree units.The Miller index labels are unitless integers. Successful indexingindicates that the sample is composed of one crystalline phase and istherefore not a mixture of crystalline phases.

IR spectroscopy, particularly FT-IR, is another technique that may beused to characterize solid forms together with or separately from X-raypowder diffraction. In an IR spectrum, absorbed light is plotted on thex-axis of a graph in the units of “wavenumber” (cm⁻¹), with intensity onthe y-axis. Variation in the position of IR peaks also exists and may bedue to sample conditions as well as data collection and processing. Thetypical variability in IR spectra reported herein is on the order ofplus or minus 2.0 cm⁻¹. Thus, the use of the word “about” whenreferencing IR peaks is meant to include this variability and all IRpeaks disclosed herein are intended to be reported with suchvariability.

Thermal methods are another typical technique to characterize solidforms. Different polymorphs of the same compound often melt at differenttemperatures. Thus, the melting point of a polymorph, as measured bymethods such as capillary melting point, DSC, and hot stage microscopy,alone or in combination with techniques such as X-ray powderdiffraction, IR spectroscopy, including FT-IR, or both, may be used tocharacterize polymorphs or other solid forms.

As with any analytical technique, melting point determinations are alsosubject to variability. Common sources of variability, in addition toinstrumental variability, are due to colligative properties such as thepresence of other solid forms or other impurities within a sample whosemelting point is being measured.

As used herein, the terms “treat,” “treating,” “treatment” andgrammatical variations thereof mean subjecting an individual subject toa protocol, regimen, process or remedy, in which it is desired to obtaina physiologic response or outcome in that subject, e.g., a patient. Inparticular, the methods and compositions of the present invention may beused to slow the development of disease symptoms or delay the onset ofthe disease or condition, or halt the progression of diseasedevelopment. However, because every treated subject may not respond to aparticular treatment protocol, regimen, process or remedy, treating doesnot require that the desired physiologic response or outcome be achievedin each and every subject or subject population, e.g., patientpopulation. Accordingly, a given subject or subject population, e.g.,patient population may fail to respond or respond inadequately totreatment.

As used herein, the terms “ameliorate”, “ameliorating” and grammaticalvariations thereof mean to decrease the severity of the symptoms of adisease in a subject.

As used herein, a “subject” is a mammal, preferably, a human. Inaddition to humans, categories of mammals within the scope of thepresent invention include, for example, farm animals, domestic animals,laboratory animals, etc. Some examples of farm animals include cows,pigs, horses, goats, etc. Some examples of domestic animals includedogs, cats, etc. Some examples of laboratory animals include primates,rats, mice, rabbits, guinea pigs, etc.

Cancers include both solid and hemotologic cancers. Non-limitingexamples of solid cancers include adrenocortical carcinoma, anal cancer,bladder cancer, bone cancer (such as osteosarcoma), brain cancer, breastcancer, carcinoid cancer, carcinoma, cervical cancer, colon cancer,endometrial cancer, esophageal cancer, extrahepatic bile duct cancer,Ewing family of cancers, extracranial germ cell cancer, eye cancer,gallbladder cancer, gastric cancer, germ cell tumor, gestationaltrophoblastic tumor, head and neck cancer, hypopharyngeal cancer, isletcell carcinoma, kidney cancer, large intestine cancer, laryngeal cancer,leukemia, lip and oral cavity cancer, liver cancer, lung cancer,lymphoma, malignant mesothelioma, Merkel cell carcinoma, mycosisfungoides, myelodysplastic syndrome, myeloproliferative disorders,nasopharyngeal cancer, neuroblastoma, oral cancer, oropharyngeal cancer,osteosarcoma, ovarian epithelial cancer, ovarian germ cell cancer,pancreatic cancer, paranasal sinus and nasal cavity cancer, parathyroidcancer, penile cancer, pituitary cancer, plasma cell neoplasm, prostatecancer, rhabdomyosarcoma, rectal cancer, renal cell cancer, transitionalcell cancer of the renal pelvis and ureter, salivary gland cancer,Sézary syndrome, skin cancers (such as cutaneous t-cell lymphoma,Kaposi's sarcoma, mast cell tumor, and melanoma), small intestinecancer, soft tissue sarcoma, stomach cancer, testicular cancer, thymoma,thyroid cancer, urethral cancer, uterine cancer, vaginal cancer, vulvarcancer, and Wilms' tumor.

Examples of hematologic cancers include, but are not limited to,leukemias, such as adult/childhood acute lymphoblastic leukemia,adult/childhood acute myeloid leukemia, chronic lymphocytic leukemia,chronic myelogenous leukemia, and hairy cell leukemia, lymphomas, suchas AIDS-related lymphoma, cutaneous T-cell lymphoma, adult/childhoodHodgkin lymphoma, mycosis fungoides, adult/childhood non-Hodgkinlymphoma, primary central nervous system lymphoma, Sézary syndrome,cutaneous T-cell lymphoma, and Waldenstrom macroglobulinemia, as well asother proliferative disorders such as chronic myeloproliferativedisorders, Langerhans cell histiocytosis, multiple myeloma/plasma cellneoplasm, myelodysplastic syndromes, andmyelodysplastic/myeloproliferative neoplasms. A preferred set of cancersthat may be treated according to the present invention includeneuroblastoma, leukemia, lymphoma, liver cancer, lung cancer, skincancer, testicular cancer, and thyroid cancer. Preferably, the cancer ismelanoma.

The methods of the present invention may optionally further includeadministering to the subject at least one additional therapeutic agenteffective for treating or ameliorating the effects of the cancer. Theadditional therapeutic agent may be selected from the group consistingof an antibody or fragment thereof, a chemotherapeutic agent, animmunotherapeutic agent, a radionuclide, a photoactive therapeuticagent, a radiosensitizing agent, and combinations thereof.

The malonate salt form of the present invention and the anti-canceragent(s) used in the co-treatment therapy may be administered to thesubject, either simultaneously or at different times, as deemed mostappropriate. If the malonate salt form of the present invention and theother anti-cancer agent(s) are administered at different times, forexample, by serial administration, then the malonate salt form of thepresent invention may be administered to the subject before the otheranti-cancer agent. Alternatively, the other anti-cancer agent(s) may beadministered to the subject before malonate salt form.

As used herein, an “antibody” encompasses naturally occurringimmunoglobulins as well as non-naturally occurring immunoglobulins,including, for example, single chain antibodies, chimeric antibodies(e.g., humanized murine antibodies), and heteroconjugate antibodies(e.g., bispecific antibodies). Fragments of antibodies include thosethat bind antigen, (e.g., Fab′, F(ab′)₂, Fab, Fv, and rIgG). See also,e.g., Pierce Catalog and Handbook, 1994-1995 (Pierce Chemical Co.,Rockford, III.); Kuby, J., Immunology, 3rd Ed., W.H. Freeman & Co., NewYork (1998). The term antibody also includes bivalent or bispecificmolecules, diabodies, triabodies, and tetrabodies. The term “antibody”further includes both polyclonal and monoclonal antibodies.

Examples of therapeutic antibodies that may be used in the presentinvention include rituximab (Rituxan), Cetuximab (Erbitux), bevacizumab(Avastin), and Ibritumomab (Zevalin).

As used herein, “chemotherapeutic agent” means any therapeutic agentthat is compatible with the malonate salt form treatment of the presentinvention and that uses cytotoxic and/or cytostatic agents againstcancer cells or cells that are associated with or support cancer cells.In a preferred embodiment, the chemotherapeutic agent is an agentselected from the group consisting of an anti-metabolite, a microtubuleinhibitor, a DNA damaging agent, an antibiotic, an anti-angiogenesisagent, a vascular disrupting agent, a molecularly targeted agent, andcombinations thereof.

As used herein, an “anti-metabolite” is a substance that reduces orinhibits a cell's use of a chemical that is part of normal metabolism.Non-limiting examples of anti-metabolite agents or analogs thereofaccording to the present invention include antifolates, purineinhibitors, pyrimidine inhibitors, and combinations thereof.

As used herein, an “antifolate” is a substance that alters, reduces, orinhibits the use of folic acid (vitamin B9) by cells. Non-limitingexamples of antifolates include methotrexate (DuraMed Pharmaceuticals,Inc.), pemetrexed (Eli Lilly), pralatrexate (Spectrum Pharmaceuticals),am inopterin (Sigma Aldrich), pharmaceutically acceptable salts thereof,and combinations thereof.

As used herein, a “purine” is a compound that contains a fusedsix-membered and a five-membered nitrogen-containing ring. Non-limitingexamples of purines that are important for cellular metabolism includeadenine, guanine, hypoxanthine, and xanthine. A “purine inhibitor” is asubstance that alters, reduces or suppresses the production of a purineor the use of a purine by a cell. Non-limiting examples of purineinhibitors include methotrexate (DuraMed Pharmaceuticals, Inc.),pemetrexed (Eli Lilly), hydroxyurea (Bristol-Myers Squibb),2-mercaptopurine (Sigma-Aldrich), 6-mercaptopurine (Sigma-Aldrich),fludarabine (Ben Venue Laboratories), clofarabine (Genzyme Corp.),nelarabine (GlaxoSmithKline), pralatrexate (Spectrum Pharmaceuticals),6-thioguanine (Gate Pharmaceuticals), forodesine (BioCrystPharmaceuticals), pentostatin (Bedford Laboratories), sapacitabine(Cyclacel Pharmaceuticals, Inc.), am inopterin (Sigma Aldrich),azathioprine (GlaxoSmithKline), pharmaceutically acceptable saltsthereof, and combinations thereof.

As used herein, a “pyrimidine” is a compound that contains asix-membered nitrogen-containing ring. Non-limiting examples ofpyrimidines that are important for cellular metabolism include uracil,thymine, cytosine, and orotic acid. A “pyrimidine inhibitor” is asubstance that alters, reduces, or suppresses the production of apyrimidine or the use of a pyrimidine by the a cell. Non-limitingexamples of pyrimidine inhibitors include 5-fluorouracil (TocrisBioscience), tegafur (LGM Pharma), capecitabine (Xeloda) (Roche),cladribine (LGM Pharma), gemcitabine (Eli Lilly), cytarabine (BedfordLaboratories), decitabine (Eisai Inc.), floxuridine (BedfordLaboratories), 5-azacytidine (Pharm ion Pharmaceuticals), doxifluridine(Cayman Chemicals), thiarabine (Access Pharmaceuticals), troxacitabine(SGX Pharmaceuticals), raltitrexed (AstraZeneca), carmofur (Santa CruzBiotechnology, Inc.), 6-azauracil (MP Biomedicals, LLC),pharmaceutically acceptable salts thereof, and combinations thereof.

In a preferred aspect of the present invention, the anti-metaboliteagent is selected from the group consisting of 5-fluorouracil (TocrisBioscience), tegafur (LGM Pharma), capecitabine (Xeloda) (Roche),cladribine (LGM Pharma), methotrexate (DuraMed Pharmaceuticals, Inc.),pemetrexed (Eli Lilly), hydroxyurea (Bristol-Myers Squibb),2-mercaptopurine (Sigma-Aldrich), 6-mercaptopurine (Sigma-Aldrich),fludarabine (Ben Venue Laboratories), gemcitabine (Eli Lilly),clofarabine (Genzyme Corp.), cytarabine (Bedford Laboratories),decitabine (Eisai Inc.), floxuridine (Bedford Laboratories), nelarabine(GlaxoSmithKline), pralatrexate (Spectrum Pharmaceuticals),6-thioguanine (Gate Pharmaceuticals), 5-azacytidine (Pharm ionPharmaceuticals), doxifluridine (Cayman Chemicals), forodesine (BioCrystPharmaceuticals), pentostatin (Bedford Laboratories), sapacitabine(Cyclacel Pharmaceuticals, Inc.), thiarabine (Access Pharmaceuticals),troxacitabine (SGX Pharmaceuticals), raltitrexed (AstraZeneca),aminopterin (Sigma Aldrich), carmofur (Santa Cruz Biotechnology, Inc.),azathioprine (GlaxoSmithKline), 6-azauracil (MP Biomedicals, LLC),pharmaceutically acceptable salts thereof, and combinations thereof.

As used herein, a “microtubule inhibitor” is a substance that disruptsthe functioning of a microtubule, such as the polymerization or thedepolymerization of individual microtubule units. In one aspect of thepresent invention, the microtubule inhibitor may be selected from thegroup consisting of a microtubule-destabilizing agent, amicrotubule-stabilizing agent, and combinations thereof. A microtubuleinhibitor of the present invention may also be selected from the groupconsisting of a taxane, a vinca alkaloid, an epothilone, andcombinations thereof. Non-limiting examples of microtubule inhibitorsaccording to the present invention include BT-062 (Biotest), HMN-214 (D.Western Therapeutics), eribulin mesylate (Eisai), vindesine (Eli Lilly),EC-1069 (Endocyte), EC-1456 (Endocyte), EC-531 (Endocyte), vintafolide(Endocyte), 2-methoxyestradiol (EntreMed), GTx-230 (GTx), trastuzumabemtansine (Hoffmann-La Roche), crolibulin (Immune Pharmaceuticals),D1302A-maytansinoid conjugates (ImmunoGen), IMGN-529 (ImmunoGen),lorvotuzumab mertansine (ImmunoGen), SAR-3419 (ImmunoGen), SAR-566658(ImmunoGen), IMP-03138 (Impact Therapeutics), topotecan/vincristinecombinations (LipoCure), BPH-8 (Molecular Discovery Systems),fosbretabulin tromethamine (OXiGENE), estramustine phosphate sodium(Pfizer), vincristine (Pierre Fabre), vinflunine (Pierre Fabre),vinorelbine (Pierre Fabre), RX-21101 (Rexahn), cabazitaxel (Sanofi),STA-9584 (Synta Pharmaceuticals), vinblastine, epothilone A, patupilone(Novartis), ixabepilone (Bristol-Myers Squibb), Epothilone D (KosanBiosciences), paclitaxel (Bristol-Myers Squibb), docetaxel(Sanofi-Aventis), HAI abraxane, DJ-927 (Daiichi Sankyo), discodermolide(CAS No: 127943-53-7), eleutherobin (CAS No.: 174545-76-7),pharmaceutically acceptable salts thereof, and combinations thereof.

DNA damaging agents of the present invention include, but are notlimited to, alkylating agents, platinum-based agents, intercalatingagents, and inhibitors of DNA replication.

As used herein, an “alkylating agent” is a substance that adds one ormore alkyl groups (C_(n)H_(m), where n and m are integers) to a nucleicacid. In the present invention, an alkylating agent is selected from thegroup consisting of nitrogen mustards, nitrosoureas, alkyl sulfonates,triazines, ethylenimines, and combinations thereof. Non-limitingexamples of nitrogen mustards include mechlorethamine (Lundbeck),chlorambucil (GlaxoSmithKline), cyclophosphamide (Mead Johnson Co.),bendamustine (Astellas), ifosfamide (Baxter International), melphalan(Ligand), melphalan flufenamide (Oncopeptides), and pharmaceuticallyacceptable salts thereof. Non-limiting examples of nitrosoureas includestreptozocin (Teva), carmustine (Eisai), lomustine (Sanofi), andpharmaceutically acceptable salts thereof. Non-limiting examples ofalkyl sulfonates include busulfan (Jazz Pharmaceuticals) andpharmaceutically acceptable salts thereof. Non-limiting examples oftriazines include dacarbazine (Bayer), temozolomide (Cancer ResearchTechnology), and pharmaceutically acceptable salts thereof. Non-limitingexamples of ethylenimines include thiotepa (Bedford Laboratories),altretamine (MGI Pharma), and pharmaceutically acceptable salts thereof.Other alkylating agents include ProLindac (Access), Ac-225 BC-8(Actinium Pharmaceuticals), ALF-2111 (Alfact Innovation), trofosfamide(Baxter International), MDX-1203 (Bristol-Myers Squibb),thioureidobutyronitrile (CellCeutix), mitobronitol (Chinoin), mitolactol(Chinoin), nimustine (Daiichi Sankyo), glufosfamide (EleisonPharmaceuticals), HuMax-TAC and PBD ADC combinations (Genmab), BP-C1(Meabco), treosulfan (Medac), nifurtimox (Metronomx), improsulfantosilate (Mitsubishi tanabe Pharma), ranimustine (Mitsubishi tanabePharma), ND-01 (NanoCarrier), HH-1 (Nordic Nanovector), 22P1G cells andifosfamide combinations (Nuvilex), estramustine phosphate (Pfizer),prednimustine (Pfizer), lurbinectedin (PharmaMar), trabectedin(PharmaMar), altreatamine (Sanofi), SGN-CD33A (Seattle Genetics),fotemustine (Servier), nedaplatin (Shionogi), heptaplatin (Sk Holdings),apaziquone (Spectrum Pharmaceuticals), SG-2000 (Spirogen), TLK-58747(Telik), laromustine (Vion Pharmaceuticals), procarbazine (AlkemLaboratories Ltd.), and pharmaceutically acceptable salts thereof.

As used herein, a “platinum-based agent” is an anti-cancer substancethat contains the metal platinum and analogs of such substances. Theplatinum may be in any oxidation state. Platinum-based agents of thepresent invention include, but are not limited to,1,2-diaminocyclohexane (DACH) derivatives, phenanthroimidazole Pt(II)complexes, platiunum IV compounds, bi- and tri-nuclear platinumcompounds, demethylcantharidin-integrated platinum complexes,platinum-conjugated compounds, cisplatin nanoparticles and polymermicelles, sterically hindered platinum complexes, oxaliplatin(Debiopharm), satraplatin (Johnson Matthey), BBR3464 (NovuspharmaS.p.A.), ZD0473 (Astra Zeneca), cisplatin (Nippon Kayaku), JM-11(Johnson Matthey), PAD (cis-dichlorobiscyclopentylamine platinum (II)),MBA ((trans-1,2-diam inocyclohexane) bisbromoacetato platinum (II)), PHM((1,2-Cyclohexanediamine) malonato platinum (II)), SHP((1,2-Cyclohexanediamine) sulphato platinum (II)), neo-PHM((trans-R,R-1,2-Cyclohexanediamine) malonato platinum (II)), neo-SHP((trans-R,R-1,2-Cyclohexanediamine)sulphato platinum (II)),JM-82(Johnson Matthey), PYP ((1,2-Cyclohexanediamine) bispyruvatoplatinum (II)), PHIC ((1,2-Cyclohexanediamine) isocitrato platinum(II)), TRK-710 ((trans-R,R-1,2-cyclohexanediamine)[3-Acetyl-5-methyl-2,4(3H,5H)-furandionato] platinum (II)), BOP((1,2-Cyclooctanediamine) bisbromoacetato platinum (II)), JM-40 (JohnsonMatthey), enloplatin (UnionPharma), zeniplatin (LGM Pharma), CI-973(Parke-Davis), lobaplatin (Zentaris AG/Hainan Tianwang InternationalPharmaceutical), cycloplatam (LGM Pharma), WA2114R(miboplatin/lobaplatin) (Chembest Research Laboratories, Ltd.),heptaplatin (SKI2053R) (SK Chemicals), TNO-6 (spiroplatin) (HaihangIndustry Co., Ltd.), ormaplatin (tetraplatin) (LGM Pharma), JM-9(iproplatin) (Johnson Matthey), BBR3610 (Novuspharma S.p.A.), BBR3005(Novuspharma S.p.A.), BBR3571 (Novuspharma S.p.A.), BBR3537 (NovuspharmaS.p.A.), aroplatin (L-NDDP) (BOC Sciences), Pt-ACRAMTU ({[Pt(en)CI(ACRAMTU-S)](NO3)2 (en=ethane-1,2-diamine,ACRAMTU=1-[2-(acridin-9-ylamino)ethyl]-1,3-dimethylthiourea)}),cisplatin-loaded liposomes (LiPlasomes), SPI-077 (Alza), lipoplatin(Regulon), lipoxal (Regulon), carboplatin (Johnson Matthey), nedaplatin(Shionogi Seiyaku), miriplatin hydrate (Dainippon Sumitomo Pharma),ormaplatin (LGM Pharma), enloplatin (Lederle Laboratories), CI973(Parke-Davis), PEGylated cisplatin, PEGylated carboplatin, PEGylatedoxaliplatin, transplatin (trans-diamminedichloroplatinum(II);mixedZ:trans-[PtCl₂{Z—HN═C(OMe)Me}(NH₃)]), CD-37 (estradiol-platinum(II)hybrid molecule), picoplatin (Poniard Pharmaceuticals),

AH44 (Komeda et al., 2006; Harris et al., 2005; Qu et al., 2004),triplatinNC (Harris et al., 2005; Qu et al., 2004), ProLindac (Access),pharmaceutically acceptable salts thereof, and combinations thereof.

As used herein, an “intercalating agent” includes, but is not limitedto, doxorubicin (Adriamycin), daunorubicin, idarubicin, mitoxantrone,pharmaceutically acceptable salts thereof, prodrugs, and combinationsthereof.

Non-limiting examples of inhibitors of DNA replication include, but arenot limited to topoisomerase inhibitors. As used herein, a“topoisomerase inhibitor” is a substance that decreases the expressionor the activity of a topoisomerase. The topoisomerase inhibitorsaccording to the present invention may inhibit topoisomerase I,topoisomerase II, or both topoisomerase I and topoisomerase II.Non-limiting examples of topoisomerase I inhibitors according to thepresent invention include irinotecan (Alchemic), APH-0804 (Aphios),camptothecin (Aphios), cositecan (BioNumerik), topotecan(GlaxoSmithKline), belotecan hydrochloride (Chon Kun Dang), firtecanpegol (Enzon), HN-30181A (Hanmi), hRS7-SN-38 (Immunomedics),labetuzumab-SN-38 (Immunomedics), etirinotecan pegol (NektarTherapeutics), NK-012 (Nippon Kayaku), SER-203 (Serina Therapeutics),simmitecan hydrochloride prodrug (Shanghai HaiHe Pharmaceuticals),gimatecan (Sigma-Tau), namitecan (Sigma-Tau), SN-38 (Supratek Pharma),TLC-388 hydrochloride (Taiwan Liposome Company), lamellarin D(PharmaMar), pharmaceutically acceptable salts thereof, and combinationsthereof. Non-limiting examples of inhibitors of topoisomerase type IIaccording to the present invention include Adva-27a (Advanomics),zoptarelin doxorubicin (Aeterna Zentaris), valrubicin (AnthraPharmaceuticals), razoxane (AstraZeneca), doxorubicin (AvenaTherapeutics), amsacrine (Bristol-Myers Squibb), etoposide phosphate(Bristol-Myers Squibb), etoposide (Novartis), dexrazoxane (CancerResearch Technology), cytarabine/daunorubicin combination (CelatorPharmaceuticals), CAP7.1 (CellAct Pharma), aldoxorubicin (CytRx),amrubicin hydrochloride (Dainippon Sumitomo Pharma), vosaroxin(Dainippon Sumitomo Pharma), daunorubicin (Gilead Sciences),milatuzumab/doxorubicin combination (Immunomedics), aclarubicin (KyowaHakko Kirin), mitoxantrone (Meda), pirarubicin (Meiji), epirubicin(Pfizer), teniposide (Novartis), F-14512 (Pierre Fabre), elliptiniumacetate (Sanofi), zorubicin (Sanofi), dexrazoxane (TopoTarget),sobuzoxane (Zenyaku Kogyo), idarubicin (Pfizer), HU-331 (CaymanChemical), aurintricarboxylic acid (Sigma Aldrich), pharmaceuticallyacceptable salts thereof, and combinations thereof.

Chemotherapeutic antibiotics according to the present invention include,but are not limited to, actinomycin, anthracyclines, valrubicin,epirubicin, bleomycin, plicamycin, mitomycin, pharmaceuticallyacceptable salts thereof, prodrugs, and combinations thereof.

As used herein, the term “anti-angiogenesis agent” means any compoundthat prevents or delays nascent blood vessel formation from existingvessels. In the present invention, examples of anti-angiogenesis agentsinclude, but are not limited to, pegaptanib, ranibizumab, bevacizumab(avastin), carboxyamidotriazole, TNP-470, CM101, IFN-α, IL-12, plateletfactor 4, suramin, SU5416, thrombospondin, VEGFR antagonists,angiostatic steroids and heparin, cartilage-derived angiogenesisinhibitory factor, matrix metalloproteinase inhibitors, angiostatin,endostatin, 2-methoxyestradiol, tecogalan, prolactin, α_(v)β₃inhibitors, linomide, VEGF-Trap, am inosterols, cortisone, tyrosinekinase inhibitors, anti-angiogenic siRNA, inhibitors of the complementsystem, vascular disrupting agents, and combinations thereof.Preferably, the anti-angiogenesis agent is bevacizumab.

VEGFR antagonists of the present invention include, but are not limitedto, pazopanib, regorafenib, lenvatinib, sorafenib, sunitinib, axitinib,vandetanib, cabozantinib, vatalanib, semaxanib, ZD6474, SU6668,AG-013736, AZD2171, AEE788, MF1/MC-18F1, DC101/IMC-1C11, ramucirumab,and motesanib. VEGFR antagonists may also include, VEGF inhibitors suchas bevacizumab, aflibercept, 2C3, r84, VEGF-Trap, and ranibizumab.

Angiostatic steroids of the present invention include any steroid thatinhibits, attenuates, prevents angiogenesis or neovascularization, orcauses regression of pathological vascularization. Angiostatic steroidsof the present invention include those disclosed in European PatentApplication Serial No. EP1236471 A2, as well as those 20-substitutedsteroids disclosed in U.S. Pat. No. 4,599,331, those 21-hydroxy steroidsdisclosed in U.S. Pat. No. 4,771,042, those C11-functionalized steroidsdisclosed in International Application Serial No. WO 1987/02672,6α-fluoro17α,21-dihydroxy-16α-methylpregna-4,9(11)-diene-3,20-dione21-acetate,6α-fluoro-17α,21-dihydroxy-16β-methylpregna-4,9(11)-diene-3,20-dione,6α-fluoro-17α,21-dihydroxy-16β-methylpregna-4,9(11)-diene-3,20-dione21-phosphonooxy and pharmaceutically acceptable salts thereof,hydrocortisone, tetrahydrocortisol, 17α-hydroxy-progesterone,11α-epihydrocortisone, cortexolone, corticosterone,desoxycorticosterone, dexamethasone, cortisone 21-acetate,hydrocortisone 21-phosphate, 17α-hydroxy-6α-methylpregn-4-ene-3,20-dione17-acetate,6α-fluoro-17α,21-dihydroxy-16α-methylpregna-4,9(11)-diene-3,20-dione,and Δ9(11)-etianic esters, all disclosed in International ApplicationSerial No. WO 1990/015816 A1.

Cartilage-derived angiogenesis inhibitor factors include, but are notlimited to, peptide troponin and chondromodulin I.

Matrix metalloproteinase inhibitors of the present invention include,but are not limited to, succinyl hydroxamates such as marimastat andSC903, sulphonamide hydroxamates such as CGS27023A, phosphinamidehydroxamates, carboxylate inhibitors such as BAY12-9566, thiolinhibitors such as Compound B, aminomethyl benzimidazole analogues,peptides such as regasepin, and tetracyclines such as minocycline.

αvβ3 inhibitors include, but are not limited to, IS20I, P11 peptide, EMD85189, and 66203, RGD peptide, RGD mimetics such as S 36578-2,echistatin, antibodies or antibody fragments against αvβ3 integrin suchas Vitaxin, which targets the extracellular domain of the dimer,cilengitide, and peptidomimetics such as S247.

Anti-angiogenic siRNAs include, but are not limited to, siRNAs targetingmRNAs that are upregulated during angiogenesis, optionally PEGylatedsiRNAs targeting VEGF or VEGFR mRNAs, and siRNAs targeting UPR (unfoldedprotein response)-IRE1α, XBP-1, and ATF6 mRNAs. Additionally, it hasbeen shown that siRNAs that are, at minimum, 21 nucleotides in length,regardless of targeting sequence, suppress neovascularization (Kleinman,et al., 2008) and may be included in the anti-angiogenic siRNAs of thepresent invention.

Inhibitors of the complement system include, but are not limited to,modified native complement components such as soluble complementreceptor type 1, soluble complement receptor type 1 lacking longhomologous repeat-A, soluble Complement Receptor Type 1-Sialyl Lewisx,complement receptor type 2, soluble decay accelerating factor, solublemembrane cofactor protein, soluble CD59, decay accelerating factor-CD59hybrid, membrane cofactor protein-decay accelerating factor hybrid, C1inhibitor, and C1q receptor, complement-inhibitory antibodies such asanti-C5 monoclonal antibody and anti-C5 single chain Fv, syntheticinhibitors of complement activation such as antagonistic peptides andanalogs targeting C5a receptor, and naturally occurring compounds thatblock complement activation such as heparin and relatedglycosaminoglycan compounds. Additional inhibitors of the complementsystem are disclosed by Makrides (Makrides, 1998).

As used herein, the term “vascular disrupting agent” means any compoundthat targets existing vasculature, e.g. tumor vasculature, damages ordestroys said vasculature, and/or causes central tumor necrosis. In thepresent invention, examples of vascular disrupting agents include, butare not limited to, ABT-751 (Abbott), AVE8062 (Aventis), BCN105(Bionomics), BMXAA (Antisoma), CA-4-P (OxiGene), CA-1-P (OxiGene),CYT997 (Cytopia), MPC-6827 (Myriad Pharmaceuticals), MN-029(MediciNova), NPI-2358 (Nereus), Oxi4503 (Oxigene), TZT-1027 (DaichiPharmaceuticals), ZD6126 (AstraZeneca and Angiogene), pharmaceuticallyacceptable salts thereof, and combinations thereof.

As used herein, a “molecularly targeted agent” is a substance thatinterferes with the function of a single molecule or group of molecules,preferably those that are involved in tumor growth and progression, whenadministered to a subject. Non-limiting examples of molecularly targetedagents of the present invention include signal transduction inhibitors,modulators of gene expression and other cellular functions, immunesystem modulators, antibody-drug conjugates (ADCs), and combinationsthereof.

As used herein, a “signal transduction inhibitor” is a substance thatdisrupts communication between cells, such as when an extracellularsignaling molecule activates a cell surface receptor. Non-limitingexamples of signal transduction inhibitors of the present inventioninclude anaplastic lymphoma kinase (ALK) inhibitors, B-Raf inhibitors,epidermal growth factor inhibitors (EGFRi), ERK inhibitors, Janus kinaseinhibitors, MEK inhibitors, mammalian target of rapamycin (mTor)inhibitors, phosphoinositide 3-kinase inhibitors (PI3Ki), and Rasinhibitors.

As used herein, an “anaplastic lymphoma kinase (ALK) inhibitor” is asubstance that (i) directly interacts with ALK, e.g., by binding to ALKand (ii) decreases the expression or the activity of ALK. Non-limitingexamples of anaplastic lymphoma kinase (ALK) inhibitors of the presentinvention include crizotinib (Pfizer, New York, N.Y.), CH5424802 (ChugaiPharmaceutical Co., Tokyo, Japan), GSK1838705 (GlaxoSmithKline, UnitedKingdom), Chugai 13d (Chugai Pharmaceutical Co., Tokyo, Japan), CEP28122(Teva Pharmaceutical Industries, Ltd., Israel), AP26113 (AriadPharmaceuticals, Cambridge, Mass.), Cephalon 30 (Teva PharmaceuticalIndustries, Ltd., Israel), X-396 (Xcovery, Inc., West Palm Beach, Fla.),Amgen 36 (Amgen Pharmaceuticals, Thousand Oaks, Calif.), ASP3026(Astellas Pharma US, Inc., Northbrook, Ill.), and Amgen 49 (AmgenPharmaceuticals, Thousand Oaks, Calif.), pharmaceutically acceptablesalts thereof, and combinations thereof.

As used herein, a “B-Raf inhibitor” of the present invention is asubstance that (i) directly interacts with B-Raf, e.g., by binding toB-Raf and (ii) decreases the expression or the activity of B-Raf. B-Rafinhibitors may be classified into two types by their respective bindingmodes. As used herein, “Type 1” B-Raf inhibitors are those inhibitorsthat target the ATP binding sites of the kinase in its activeconformation. “Type 2” B-Raf inhibitors are those inhibitors thatpreferentially bind to an inactive conformation of the kinase.Non-limiting examples of Type 1 B-Raf inhibitors of the presentinvention include:

dabrafenib (GlaxoSmithKline), GDC-0879 (Genentech), L-779450 B-Raf(Merck), PLX3202 (Plexxikon), PLX4720 (Plexxikon), SB-590885(GlaxoSmithKline), SB-699393 (GlaxoSmithKline), vemurafenib (Plexxikon),pharmaceutically acceptable salts thereof, and combinations thereof.Preferably, the type 1 RAF inhibitor is dabrafenib or a pharmaceuticallyacceptable salt thereof.

Non-limiting examples of Type 2 B-Raf inhibitors of the presentinvention include:

Sorafenib (Onyx Pharmaceuticals), ZM-336372 (AstraZeneca),pharmaceutically acceptable salts thereof, and combinations thereof

Other B-Raf inhibitors include, without limitation, AAL881 (Novartis);AB-024 (Ambit Biosciences), ARQ-736 (ArQule), ARQ-761 (ArQule), AZ628(Axon Medchem BV), BeiGene-283 (BeiGene), BUB-024 (MLN 2480) (Sunesis &Takeda), b raf inhibitor (Sareum), BRAF kinase inhibitor (SelexagenTherapeutics), BRAF siRNA 313 (tacaccagcaagctagatgca) and 253(cctatcgttagagtcttcctg) (Liu et al., 2007), CTT239065 (Institute ofCancer Research), DP-4978 (Deciphera Pharmaceuticals), HM-95573 (Hanmi),GW 5074 (Sigma Aldrich), ISIS 5132 (Novartis), LErafAON (NeoPharm,Inc.), LBT613 (Novartis), LGX 818 (Novartis), pazopanib(GlaxoSmithKline), PLX5568 (Plexxikon), RAF-265 (Novartis), RAF-365(Novartis), regorafenib (Bayer Healthcare Pharmaceuticals, Inc.), RO5126766 (Hoffmann-La Roche), TAK 632 (Takeda), TL-241 (Teligene), XL-281(Exelixis), pharmaceutically acceptable salts thereof, and combinationsthereof.

As used herein, an “EGFR inhibitor” is a substance that (i) directlyinteracts with EGFR, e.g. by binding to EGFR and (ii) decreases theexpression or the activity of EGFR. Non-limiting examples of EGFRinhibitors according to the present invention include (+)-Aeroplysinin-1(CAS #28656-91-9), 3-(4-Isopropylbenzylidenyl)-indolin-2-one, ABT-806(Life Science Pharmaceuticals), AC-480 (Bristol-Myers Squibb), afatinib(Boehringer Ingelheim), AG 1478 (CAS #153436-53-4), AG 494 (CAS#133550-35-3), AG 555 (CAS #133550-34-2), AG 556 (CAS #133550-41-1), AG825 (CAS #149092-50-2), AG-490 (CAS #134036-52-5), antroquinonol (GoldenBiotechnology), AP-26113 (Ariad), ARRY334543 (CAS #845272-21-1), AST1306 (CAS #897383-62-9), AVL-301 (Celgene), AZD8931 (CAS #848942-61-0),BIBU 1361 (CAS #793726-84-8), BIBX 1382 (CAS #196612-93-8), BMS-690514(Bristol-Myers Squibb), BPIQ-I (CAS #174709-30-9), Canertinib (Pfizer),cetuximab (Actavis), cipatinib (Jiangsu Hengrui Medicine), CL-387,785(Santa Cruz Biotech), compound 56 (CAS #171745-13-4), CTX-023 (CytomXTherapeutics), CUDC-101 (Curis), dacomitinib (Pfizer), DAPH (CAS#145915-58-8), daphnetin (Santa Cruz Biotech), dovitinib lactate(Novartis), EGFR Inhibitor (CAS #879127-07-8), epitinib (Hutchison ChinaMediTech), erbstatin Analog (CAS #63177-57-1), erlotinib (Astellas),gefitinib (AstraZeneca), GT-MAB 5.2-GEX (Glycotope), GW 583340 (CAS#388082-81-3), GW2974 (CAS #202272-68-2), HDS 029 (CAS #881001-19-0),Hypericin (Santa Cruz Biotech), icotinib hydrochloride (Betapharma),JNJ-26483327 (Johnson & Johnson), JNJ-28871063 (Johnson & Johnson),KD-020 (Kadmon Pharmaceuticals), lapatinib ditosylate (GlaxoSmithKline),Lavendustin A (Sigma), Lavendustin C (Sigma), LY-3016859 (Eli Lilly),MEHD-7945A (Hoffmann-La Roche), MM-151 (Merrimack), MT-062 (MedisynTechnologies), necitumumab (Eli Lilly), neratinib (Pfizer), nimotuzumab(Center of Molecular Immunology), NT-004 (NewGen Therapeutics),pantiumumab (Amgen), PD 153035 (CAS #153436-54-5), PD 161570 (CAS#192705-80-9), PD 168393, PD 174265 (CAS #216163-53-0), pirotinib(Sihuan Pharmaceutical), poziotinib (Hanmi), PP 3 (CAS #5334-30-5),PR-610 (Proacta), pyrotinib (Jiangsu Hengrui Medicine), RG-13022 (CAS#136831-48-6), rindopepimut (Celldex Therapeutics), RPI-1 (CAS#269730-03-2), S-222611 (Shionogi), TAK 285 (CAS #871026-44-7), TAS-2913(Taiho), theliatinib (Hutchison China MediTech), Tyrphostin 47(RG-50864, AG-213) (CAS #118409-60-2), Tyrphostin 51 (CAS #122520-90-5),Tyrphostin AG 1478 (CAS #175178-82-2), Tyrphostin AG 183 (CAS#126433-07-6), Tyrphostin AG 528 (CAS #133550-49-9), Tyrphostin AG 99(CAS #118409-59-9), Tyrphostin B42 (Santa Cruz Biotech), Tyrphostin B44(Santa Cruz Biotech), Tyrphostin RG 14620 (CAS #136831-49-7), vandetanib(AstraZeneca), varlitinib (Array BioPharma), vatalanib (Novartis), WZ3146 (CAS #1214265-56-1), WZ 4002 (CAS #1213269-23-8), WZ8040 (CAS#1214265-57-2), XL-647 (Exelixis), Z-650 (HEC Pharm), ZM 323881 (CAS#324077-30-7), pharmaceutically acceptable salts thereof, andcombinations thereof. Preferably, the EGFR inhibitor is selected fromthe group consisting of panitumumab, erlotinib, pharmaceuticallyacceptable salts thereof, and combinations thereof.

As noted above, the malonate salt of the present invention is an ERKinhibitor. As used herein, an “ERK inhibitor” is a substance that (i)directly interacts with ERK, including ERK1 and ERK2, e.g., by bindingto ERK and (ii) decreases the expression or the activity of an ERKprotein kinase. Therefore, inhibitors that act upstream of ERK, such asMEK inhibitors and RAF inhibitors, are not ERK inhibitors according tothe present invention. The malonate salt of the present invention may beadministered as a combination therapy together with other ERKinhibitors, which include, for example, AEZS-131 (Aeterna Zentaris),AEZS-136 (Aeterna Zentaris), SCH-722984 (Merck & Co.), SCH-772984 (Merck& Co.), SCH-900353 (MK-8353) (Merck & Co.), pharmaceutically acceptablesalts thereof, and combinations thereof.

As used herein, a “Janus kinase inhibitor” is a substance that (i)directly interacts with a Janus kinase, e.g., by binding to a Januskinase and (ii) decreases the expression or the activity of a Januskinase. Janus kinases of the present invention include Tyk2, Jak1, Jak2,and Jak3. Non-limiting examples of Janus kinase inhibitors of thepresent invention include ruxolitinib (Incyte Corporation, Wilmington,Del.), baricitinib (Incyte Corporation, Wilmington, Del.), tofacitinib(Pfizer, New York, N.Y.), VX-509 (Vertex Pharmaceuticals, Inc., Boston,Mass.), GLPG0634 (Galapagos NV, Belgium), CEP-33779 (TevaPharmaceuticals, Israel), pharmaceutically acceptable salts thereof, andcombinations thereof

As used herein, a “MEK inhibitor” is a substance that (i) directlyinteracts with MEK, e.g., by binding to MEK and (ii) decreases theexpression or the activity of MEK. Therefore, inhibitors that actupstream of MEK, such as RAS inhibitors and RAF inhibitors, are not MEKinhibitors according to the present invention. MEK inhibitors may beclassified into two types depending on whether the inhibitor competeswith ATP. As used herein, a “Type 1” MEK inhibitor is an inhibitor thatcompetes with ATP for binding to MEK. A “Type 2” MEK inhibitor is aninhibitor that does not compete with ATP for binding to MEK.Non-limiting examples of type 1 MEK inhibitors according to the presentinvention include bentamapimod (Merck KGaA), L783277 (Merck), RO092210(Roche), pharmaceutically acceptable salts thereof, and combinationsthereof. Preferably, the type 1 MEK inhibitor is RO092210 (Roche) or apharmaceutically acceptable salt thereof. Non-limiting examples of type2 MEK inhibitors according to the present invention include anthraxtoxin, lethal factor portion of anthrax toxin, ARRY-142886(6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carboxylicacid (2-hydroxy-ethoxy)-amide) (Array BioPharma), ARRY-438162 (ArrayBioPharma), AS-1940477 (Astellas), MEK162 (Array BioPharma), PD 098059(2-(2′-amino-3′-methoxyphenyl)-oxanaphthalen-4-one), PD 184352(CI-1040), PD-0325901 (Pfizer), pimasertib (Santhera Pharmaceuticals),refametinib (AstraZeneca), selumetinib (AZD6244) (AstraZeneca), TAK-733(Takeda), trametinib (Japan Tobacco), U0126 (1,4-diamino-2,3-dicyano-1,4-bis(2-aminophenylthio)butadiene) (Sigma), RDEA119(Ardea Biosciences/Bayer), pharmaceutically acceptable salts thereof,and combinations thereof. Preferably, the type 2 MEK inhibitor istrametinib or a pharmaceutically acceptable salt thereof. Other MEKinhibitors include, without limitation, antroquinonol (GoldenBiotechnology), AS-1940477 (Astellas), AS-703988 (Merck KGaA), BI-847325(Boehringer Ingelheim), E-6201 (Eisai), GDC-0623 (Hoffmann-La Roche),GDC-0973, RG422, RO4987655, RO5126766, SL327, WX-554 (Wilex), YopJpolypeptide, pharmaceutically acceptable salts thereof, and combinationsthereof.

As used herein, an “mTOR inhibitor” is a substance that (i) directlyinteracts with mTOR, e.g. by binding to mTOR and (ii) decreases theexpression or the activity of mTOR. Non-limiting examples of mTORinhibitors according to the present invention include zotarolimus(AbbVie), umirolimus (Biosensors), temsirolimus (Pfizer), sirolimus(Pfizer), sirolimus NanoCrystal (Elan Pharmaceutical Technologies),sirolimus TransDerm (TransDerm), sirolimus-PNP (Samyang), everolimus(Novartis), biolimus A9 (Biosensors), ridaforolimus (Ariad), rapamycin,TCD-10023 (Terumo), DE-109 (MacuSight), MS-R001 (MacuSight), MS-R002(MacuSight), MS-R003 (MacuSight), Perceiva (MacuSight), XL-765(Exelixis), quinacrine (Cleveland BioLabs), PKI-587 (Pfizer),PF-04691502 (Pfizer), GDC-0980 (Genentech and Piramed), dactolisib(Novartis), CC-223 (Celgene), PWT-33597 (Pathway Therapeutics), P-7170(Piramal Life Sciences), LY-3023414 (Eli Lilly), INK-128 (Takeda),GDC-0084 (Genentech), DS-7423 (Daiichi Sankyo), DS-3078 (DaiichiSankyo), CC-115 (Celgene), CBLC-137 (Cleveland BioLabs), AZD-2014(AstraZeneca), X-480 (Xcovery), X-414 (Xcovery), EC-0371 (Endocyte),VS-5584 (Verastem), PQR-401 (Piqur), PQR-316 (Piqur), PQR-311 (Piqur),PQR-309 (Piqur), PF-06465603 (Pfizer), NV-128 (Novogen), nPT-MTOR(Biotica Technology), BC-210 (Biotica Technology), WAY-600 (BioticaTechnology), WYE-354 (Biotica Technology), WYE-687 (Biotica Technology),LOR-220 (Lorus Therapeutics), HMPL-518 (Hutchison China MediTech),GNE-317 (Genentech), EC-0565 (Endocyte), CC-214 (Celgene), and ABTL-0812(Ability Pharmaceuticals).

As used herein, a “PI3K inhibitor” is a substance that decreases theexpression or the activity of phosphatidylinositol-3 kinases (PI3Ks) ordownstream proteins, such as Akt. PI3Ks, when activated, phosphorylatethe inositol ring 3′-OH group in inositol phospholipids to generate thesecond messenger phosphatidylinositol-3,4,5-trisphosphate(PI-3,4,5-P(3)). Akt interacts with a phospholipid, causing it totranslocate to the inner membrane, where it is phosphorylated andactivated. Activated Akt modulates the function of numerous substratesinvolved in the regulation of cell survival, cell cycle progression andcellular growth.

Non-limiting examples of PI3K inhibitors according to the presentinvention include A-674563 (CAS #552325-73-2), AGL 2263, AMG-319 (Amgen,Thousand Oaks, Calif.), AS-041164(5-benzo[1,3]dioxol-5-ylmethylene-thiazolidine-2,4-dione), AS-604850(5-(2,2-Difluoro-benzo[1,3]dioxol-5-ylmethylene)-thiazolidine-2,4-dione),AS-605240 (5-quinoxilin-6-methylene-1,3-thiazolidine-2,4-dione), AT7867(CAS #857531-00-1), benzimidazole series, Genentech (Roche HoldingsInc., South San Francisco, Calif.), BML-257 (CAS #32387-96-5), CAL-120(Gilead Sciences, Foster City, Calif.), CAL-129 (Gilead Sciences),CAL-130 (Gilead Sciences), CAL-253 (Gilead Sciences), CAL-263 (GileadSciences), CAS #612847-09-3, CAS #681281-88-9, CAS #75747-14-7, CAS#925681-41-0, CAS #98510-80-6, CCT128930 (CAS #885499-61-6), CH5132799(CAS #1007207-67-1), CHR-4432 (Chroma Therapeutics, Ltd., Abingdon, UK),FPA 124 (CAS #902779-59-3), GS-1101 (CAL-101) (Gilead Sciences), GSK690693 (CAS #937174-76-0), H-89 (CAS #127243-85-0), Honokiol, IC87114(Gilead Science), IPI-145 (Intellikine Inc.), KAR-4139 (KarusTherapeutics, Chilworth, UK), KAR-4141 (Karus Therapeutics), KIN-1(Karus Therapeutics), KT 5720 (CAS #108068-98-0), Miltefosine, MK-2206dihydrochloride (CAS #1032350-13-2), ML-9 (CAS #105637-50-1),Naltrindole Hydrochloride, OXY-111A (NormOxys Inc., Brighton, Mass.),perifosine, PHT-427 (CAS #1191951-57-1), PI3 kinase delta inhibitor,Merck KGaA (Merck & Co., Whitehouse Station, N.J.), PI3 kinase deltainhibitors, Genentech (Roche Holdings Inc.), PI3 kinase deltainhibitors, Incozen (Incozen Therapeutics, Pvt. Ltd., Hydrabad, India),PI3 kinase delta inhibitors-2, Incozen (Incozen Therapeutics), PI3kinase inhibitor, Roche-4 (Roche Holdings Inc.), PI3 kinase inhibitors,Roche (Roche Holdings Inc.), PI3 kinase inhibitors, Roche-5 (RocheHoldings Inc.), PI3-alpha/delta inhibitors, Pathway Therapeutics(Pathway Therapeutics Ltd., South San Francisco, Calif.), PI3-deltainhibitors, Cellzome (Cellzome AG, Heidelberg, Germany), PI3-deltainhibitors, Intellikine (Intellikine Inc., La Jolla, Calif.), PI3-deltainhibitors, Pathway Therapeutics-1 (Pathway Therapeutics Ltd.),PI3-delta inhibitors, Pathway Therapeutics-2 (Pathway TherapeuticsLtd.), PI3-delta/gamma inhibitors, Cellzome (Cellzome AG),PI3-delta/gamma inhibitors, Cellzome (Cellzome AG), PI3-delta/gammainhibitors, Intellikine (Intellikine Inc.), PI3-delta/gamma inhibitors,Intellikine (Intellikine Inc.), PI3-delta/gamma inhibitors, PathwayTherapeutics (Pathway Therapeutics Ltd.), PI3-delta/gamma inhibitors,Pathway Therapeutics (Pathway Therapeutics Ltd.), PI3-gamma inhibitorEvotec (Evotec), PI3-gamma inhibitor, Cellzome (Cellzome AG), PI3-gammainhibitors, Pathway Therapeutics (Pathway Therapeutics Ltd.), PI3Kdelta/gamma inhibitors, Intellikine-1 (Intellikine Inc.), PI3Kdelta/gamma inhibitors, Intellikine-1 (Intellikine Inc.), pictilisib(GDC-0941) (Roche Holdings Inc.), PIK-90 (CAS #677338-12-4), SC-103980(Pfizer, New York, N.Y.), SF-1126 (Semafore Pharmaceuticals,Indianapolis, Ind.), SH-5, SH-6, Tetrahydro Curcumin, TG100-115(Targegen Inc., San Diego, Calif.), Triciribine, X-339 (Xcovery, WestPalm Beach, Fla.), XL-499 (Evotech, Hamburg, Germany), pharmaceuticallyacceptable salts thereof, and combinations thereof. Preferably, theinhibitor of the PI3K/Akt pathway is pictilisib (GDC-0941) or apharmaceutically acceptable salt thereof.

As used herein, a “RAS inhibitor” is a substance that (i) directlyinteracts with RAS, e.g., by binding to RAS and (ii) decreases theexpression or the activity of RAS. Non-limiting examples of RASinhibitors according to the present invention include farnesyltransferase inhibitors (such as, e.g., tipifarnib and lonafarnib),farnesyl group-containing small molecules (such as, e.g., salirasib andTLN-4601), DCAI, as described by Maurer (Maurer, et al., 2012), Kobe0065and Kobe2602, as described by Shima (Shima, et al., 2013), and HBS 3(Patgiri, et al., 2011), and AIK-4 (Allinky), pharmaceuticallyacceptable salts thereof, and combinations thereof.

As used herein, “gene expression” is a process by which the informationfrom DNA is used in the formation of a polypeptide. A “modulator of geneexpression and other cellular functions” is a substance that affectsgene expression and other works of a cell. Non-limiting examples of suchmodulators include hormones, histone deacetylase inhibitors (HDACi), andcyclin-dependent kinase inhibitors (CDKi), and poly ADP ribosepolymerase (PARP) inhibitors.

In the present invention, a “hormone” is a substance released by cellsin one part of a body that affects cells in another part of the body.Non-limiting examples of hormones according to the present inventioninclude prostaglandins, leukotrienes, prostacyclin, thromboxane, amylin,antimullerian hormone, adiponectin, adrenocorticotropic hormone,angiotensinogen, angiotensin, vasopressin, atriopeptin, brainnatriuretic peptide, calcitonin, cholecystokinin,corticotropin-releasing hormone, encephalin, endothelin, erythropoietin,follicle-stimulating hormone, galanin, gastrin, ghrelin, glucagon,gonadotropin-releasing hormone, growth hormone-releasing hormone, humanchorionic gonadotropin, human placental lactogen, growth hormone,inhibin, insulin, somatomedin, leptin, liptropin, luteinizing hormone,melanocyte stimulating hormone, motilin, orexin, oxytocin, pancreaticpolypeptide, parathyroid hormone, prolactin, prolactin releasinghormone, relaxin, renin, secretin, somatostain, thrombopoietin,thyroid-stimulating hormone, testosterone, dehydroepiandrosterone,androstenedione, dihydrotestosterone, aldosterone, estradiol, estrone,estriol, cortisol, progesterone, calcitriol, and calcidiol.

Some compounds interfere with the activity of certain hormones or stopthe production of certain hormones. Non-limiting examples ofhormone-interfering compounds according to the present invention includetamoxifen (Nolvadex®), anastrozole (Arimidex®), letrozole (Femara®), andfulvestrant (Faslodex®). Such compounds are also within the meaning ofhormone in the present invention.

As used herein, an “HDAC inhibitor” is a substance that (i) directlyinteracts with HDAC, e.g., by binding to HDAC and (ii) decreases theexpression or the activity of HDAC. Non-limiting examples of HDACinhibitors according to the present invention include 4SC-201 (4SC AG),4SC-202 (Takeda), abexinostat (Celera), AN-1 (Titan Pharmaceuticals,Inc.), Apicidine (Merck & Co., Inc.), AR-42 (Arno Therapeutics),ARQ-700RP (ArQule), Avugane (TopoTarget AS),azelaic-1-hydroxamate-9-anilide (AAHA), belinostat (TopoTarget),butyrate (Enzo Life Sciences, Inc.), CG-1255 (Errant Gene Therapeutics,LLC), CG-1521 (Errant Gene Therapeutics, LLC), CG-200745(CrystalGenomics, Inc.), chidamide (Shenzhen Chipscreen Biosciences),CHR-3996 (Chroma Therapeutics), CRA-024781 (Pharmacyclics), CS-3158(Shenzhen Chipscreen Biosciences), CU-903 (Curis), DAC-60 (Genextra),entinostat (Bayer), hyaluronic acid butyric acid ester (HA-But), IKH-02(IkerChem), IKH-35 (IkerChem), ITF-2357 (Italfarmaco), ITF-A(Italfarmaco), JNJ-16241199 (Johnson & Johnson), KA-001 (KarusTherapeutics), KAR-3000 (Karus Therapeutics), KD-5150 (Kalypsys),KD-5170 (Kalypsys), KLYP-278 (Kalypsys), KLYP-298 (Kalypsys), KLYP-319(Kalypsys), KLYP-722 (Kalypsys), m-carboxycinnamic acid bis-hydroxamide(CBHA), MG-2856 (MethylGene), MG-3290 (MethylGene), MG-4230(MethylGene), MG-4915 (MethylGene), MG-5026 (MethylGene), MGCD-0103(MethylGene Inc.), mocetinostat (MethylGene), MS-27-275 (Schering AG),NBM-HD-1 (NatureWise), NVP-LAQ824 (Novartis), OCID-4681-S-01 (OrchidPharmaceuticals), oxamflatin ((2E)-5-[3-[(phenylsufonyl) aminolphenyl]-pent-2-en-4-ynohydroxamic acid), panobinostat (Novartis),PCI-34051 (Pharmacyclics), phenylbutyrate (Enzo Life Sciences, Inc.),pivaloyloxymethyl butyrate (AN-9, Titan Pharmaceuticals, Inc.), pivanex(Titan Pharmaceuticals, Inc.), pracinostat (SBIO), PX-117794 (TopoTargetAS), PXD-118490 (LEO-80140) (TopoTarget AS), pyroxam ide (suberoyl-3-aminopyridineam ide hydroxamic acid), resminostat (Takeda), RG-2833(RepliGen), ricolinostat (Acetylon), romidepsin (Astellas), SB-1304(S*BIO), SB-1354 (S*BIO), SB-623 (Merrion Research I Limited), SB-624(Merrion Research I Limited), SB-639 (Merrion Research I Limited),SB-939 (S*BIO), Scriptaid(N-Hydroxy-1,3-dioxo-1H-benz[de]isoquinoline-2(3H)-hexan amide), SK-7041(In2Gen/SK Chemical Co.), SK-7068 (In2Gen/SK Chemical Co.),suberoylanilide hydroxamic acid (SAHA), sulfonamide hydroxamic acid,tributyrin (Sigma Aldrich), trichostatin A (TSA) (Sigma Aldrich),valporic acid (VPA) (Sigma Aldrich), vorinostat (Zolinza), WF-27082B(Fujisawa Pharmaceutical Company, Ltd.), pharmaceutically acceptablesalts thereof, and combinations thereof. Preferably, the HDAC inhibitoris romidepsin, pharmaceutically acceptable salts thereof, andcombinations thereof.

As used herein, “CDK” is a family of protein kinases that regulate thecell cycle. Known CDKs include cdk1, cdk2, ckd3, ckd4, cdk5, cdk6, cdk7,cdk8, cdk9, cdk10, and cdk11. A “CDK inhibitor” is a substance that (i)directly interacts with CDK, e.g. by binding to CDK and (ii) decreasesthe expression or the activity of CDK. Non-limiting examples of CDKinhibitors according to the present invention include 2-Hydroxybohemine,3-ATA, 5-Iodo-Indirubin-3′-monoxime, 9-Cyanopaullone, Aloisine A,Alsterpaullone 2-Cyanoethyl, alvocidib (Sanofi), AM-5992 (Amgen),Aminopurvalanol A, Arcyriaflavin A, AT-7519 (Astex Pharmaceuticals), AZD5438 (CAS #602306-29-6), BMS-265246 (CAS #582315-72-8), BS-181 (CAS#1092443-52-1), Butyrolactone I (CAS #87414-49-1), Cdk/Crk Inhibitor(CAS #784211-09-2), Cdk1/5 Inhibitor (CAS #40254-90-8), Cdk2 InhibitorII (CAS #222035-13-4), Cdk2 Inhibitor IV, NU6140 (CAS #444723-13-1),Cdk4 Inhibitor (CAS #546102-60-7), Cdk4 Inhibitor III (CAS#265312-55-8), Cdk4/6 Inhibitor IV (CAS #359886-84-3), Cdk9 Inhibitor II(CAS #140651-18-9), CGP 74514A, CR8, CYC-065 (Cyclacel), dinaciclib(Ligand), (R)-DRF053 dihydrochloride (CAS #1056016-06-8), Fascaplysin,Flavopiridol, Hygrolidin, Indirubin, LEE-011 (Astex Pharmaceuticals),LY-2835219 (Eli Lilly), milciclib maleate (Nerviano Medical Sciences),MM-D37K (Maxwell Biotech), N9-Isopropyl-olomoucine, NSC 625987 (CAS#141992-47-4), NU2058 (CAS #161058-83-9), NU6102 (CAS #444722-95-6),Olomoucine, ON-108600 (Onconova), ON-123300 (Onconova), Oxindole I,P-1446-05 (Piramal), P-276-00 (Piramal), palbociclib (Pfizer),PHA-767491 (CAS #845714-00-3), PHA-793887 (CAS #718630-59-2), PHA-848125(CAS #802539-81-7), Purvalanol A, Purvalanol B, R547 (CAS #741713-40-6),RO-3306 (CAS #872573-93-8), Roscovitine, SB-1317 (SBIO), SCH 900776 (CAS#891494-63-6), SEL-120 (Selvita), seliciclib (Cyclacel), SNS-032 (CAS#345627-80-7), SU9516 (CAS #377090-84-1), WHI-P180 (CAS #211555-08-7),pharmaceutically acceptable salts thereof, and combinations thereof.Preferably, the CDK inhibitor is selected from the group consisting ofdinaciclib, palbociclib, pharmaceutically acceptable salts thereof, andcombinations thereof.

As used herein, a “poly ADP ribose polymerase (PARP) inhibitor” is asubstance that decreases the expression or activity of poly ADP ribosepolymerases (PARPs) or downstream proteins. Non-limiting examples ofpoly ADP ribose polymerase (PARP) inhibitors of the present inventioninclude PF01367338 (Pfizer, New York, N.Y.), olaparib (AstraZeneca,United Kingdom), iniparib (Sanofi-Aventis, Paris, France), veliparib(Abbott Laboratories, Abbott Park, Ill.), MK 4827 (Merck, White HouseStation, N.J.), CEP 9722 (Teva Pharmaceuticals, Israel), LT-673(Biomarin, San Rafael, Calif.), and BSI 401 (Sanofi-Aventis, Paris,France), pharmaceutically acceptable salts thereof, and combinationsthereof.

As used herein, “immunotherapeutic agent” means any anti-cancer agentthat is compatible with the solid forms of the present invention andthat uses a substance that alters the immune response by augmenting orreducing the ability of the immune system to produce antibodies orsensitized cells that recognize and react with the antigen thatinitiated their production. Immunotherapeutic agents may be recombinant,synthetic, or natural preparations and include cytokines,corticosteroids, cytotoxic agents, thymosin, and immunoglobulins. Someimmunotherapeutic agents are naturally present in the body, and certainof these are available in pharmacologic preparations. Examples ofimmunotherapeutic agents include, but are not limited to, granulocytecolony-stimulating factor (G-CSF), interferons, imiquimod and cellularmembrane fractions from bacteria, IL-2, IL-7, IL-12, CCL3, CCL26, CXCL7,and synthetic cytosine phosphate-guanosine (CpG).

In one preferred embodiment, the immunotherapeutic agent is an immunecheckpoint inhibitor. As used herein, an “immune checkpoint inhibitor”means a substance that blocks the activity of molecules involved inattenuating the immune response. Such molecules include, for example,cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) and programmed celldeath protein 1 (PD-1). Immune checkpoint inhibitors of the presentinvention include, but are not limited to, ipilimumab (Bristol-MyersSquibb), tremelimumab (Pfizer), MDX-1106 (Medarex, Inc.), MK3475(Merck), CT-011 (CureTech, Ltd.), AMP-224 (Amplmmune), MDX-1105(Medarex, Inc.), IMP321 (Immutep S.A.), and MGA271 (Macrogenics).

In the present invention, the term “radionuclide” means a radioactivesubstance administered to the patient, e.g., intravenously or orally,after which it penetrates via the patient's normal metabolism into thetarget organ or tissue, where it delivers local radiation for a shorttime. Examples of radionuclides include, but are not limited to, I-125,At-211, Lu-177, Cu-67, I-131, Sm-153, Re-186, P-32, Re-188, In-114m, andY-90.

In the present invention, the term “photoactive therapeutic agent” meanscompounds and compositions that become active upon exposure to light.Certain examples of photoactive therapeutic agents are disclosed, e.g.,in U.S. Patent Application Serial No. 2011/0152230 A1, “PhotoactiveMetal Nitrosyls For Blood Pressure Regulation And Cancer Therapy.”

In the present invention, the term “radiosensitizing agent” means acompound that makes tumor cells more sensitive to radiation therapy.Examples of radiosensitizing agents include misonidazole, metronidazole,tirapazamine, and trans sodium crocetinate.

In the present invention, an “effective amount” or a “therapeuticallyeffective amount” of the malonate salt form of the present invention oranother anti-cancer agent of the invention, including the pharmaceuticalcompositions containing same, is an amount of such malonate salt form orcomposition that is sufficient to effect beneficial or desired resultsas described herein when administered to a subject. Effective dosageforms, modes of administration, and dosage amounts may be determinedempirically, and making such determinations is within the skill of theart. It is understood by those skilled in the art that the dosage amountwill vary with the route of administration, the rate of excretion, theduration of the treatment, the identity of any other drugs beingadministered, the age, size, and species of subject, e.g., humanpatient, and like factors well known in the arts of medicine andveterinary medicine. In general, a suitable dose of one or more of themalonate salt form of the present invention or a pharmaceuticalcomposition according to the invention will be that amount of themalonate salt form or pharmaceutical composition, which is the lowestdose effective to produce the desired effect. The effective dose of amalonate salt form or pharmaceutical composition of the presentinvention may be administered as two, three, four, five, six or moresub-doses, administered separately at appropriate intervals throughoutthe day.

A suitable, non-limiting example of a dosage of a malonate salt form ofthe present invention or another anti-cancer agent disclosed herein isfrom about 1 mg/kg to about 2400 mg/kg per day, such as from about 1mg/kg to about 1200 mg/kg per day, 75 mg/kg per day to about 300 mg/kgper day, including from about 1 mg/kg to about 100 mg/kg per day. Otherrepresentative dosages of such agents include about 1 mg/kg, 5 mg/kg, 10mg/kg, 15 mg/kg, 20 mg/kg, 25 mg/kg, 30 mg/kg, 35 mg/kg, 40 mg/kg, 45mg/kg, 50 mg/kg, 60 mg/kg, 70 mg/kg, 75 mg/kg, 80 mg/kg, 90 mg/kg, 100mg/kg, 125 mg/kg, 150 mg/kg, 175 mg/kg, 200 mg/kg, 250 mg/kg, 300 mg/kg,400 mg/kg, 500 mg/kg, 600 mg/kg, 700 mg/kg, 800 mg/kg, 900 mg/kg, 1000mg/kg, 1100 mg/kg, 1200 mg/kg, 1300 mg/kg, 1400 mg/kg, 1500 mg/kg, 1600mg/kg, 1700 mg/kg, 1800 mg/kg, 1900 mg/kg, 2000 mg/kg, 2100 mg/kg, 2200mg/kg, and 2300 mg/kg per day. The effective dose of a malonate saltform of the present invention or other anti-cancer agents disclosedherein may be administered as two, three, four, five, six or moresub-doses, administered separately at appropriate intervals throughoutthe day.

The malonate salt form of the present invention or other anti-canceragents or pharmaceutical compositions containing same of the presentinvention may be administered in any desired and effective manner: fororal ingestion, or as an ointment or drop for local administration tothe eyes, or for parenteral or other administration in any appropriatemanner such as intraperitoneal, subcutaneous, topical, intradermal,inhalation, intrapulmonary, rectal, vaginal, sublingual, intramuscular,intravenous, intraarterial, intrathecal, or intralymphatic. Further, themalonate salt form of the present invention or other anti-cancer agentsor pharmaceutical compositions containing same of the present inventionmay be administered in conjunction with other treatments. The malonatesalt form of the present invention or other anti-cancer agents or thepharmaceutical compositions of the present invention may be encapsulatedor otherwise protected against gastric or other secretions, if desired.

The pharmaceutical compositions of the invention may comprise one ormore active ingredients, e.g., malonate salt forms of the presentinvention optionally in combination with other anti-cancer agents, inadmixture with one or more pharmaceutically-acceptable diluents orcarriers and, optionally, one or more other compounds, drugs,ingredients and/or materials. Regardless of the route of administrationselected, the agents/compounds of the present invention are formulatedinto pharmaceutically-acceptable dosage forms by conventional methodsknown to those of skill in the art. See, e.g., Remington, The Scienceand Practice of Pharmacy (21st Edition, Lippincott Williams and Wilkins,Philadelphia, Pa.).

Pharmaceutically acceptable diluents or carriers are well known in theart (see, e.g., Remington, The Science and Practice of Pharmacy (21stEdition, Lippincott Williams and Wilkins, Philadelphia, Pa.) and TheNational Formulary (American Pharmaceutical Association, Washington,D.C.)) and include sugars (e.g., lactose, sucrose, mannitol, andsorbitol), starches, cellulose preparations, calcium phosphates (e.g.,dicalcium phosphate, tricalcium phosphate and calcium hydrogenphosphate), sodium citrate, water, aqueous solutions (e.g., saline,sodium chloride injection, Ringer's injection, dextrose injection,dextrose and sodium chloride injection, lactated Ringer's injection),alcohols (e.g., ethyl alcohol, propyl alcohol, and benzyl alcohol),polyols (e.g., glycerol, propylene glycol, and polyethylene glycol),organic esters (e.g., ethyl oleate and triglycerides), biodegradablepolymers (e.g., polylactide-polyglycolide, poly(orthoesters), andpoly(anhydrides)), elastomeric matrices, liposomes, microspheres, oils(e.g., corn, germ, olive, castor, sesame, cottonseed, and groundnut),cocoa butter, waxes (e.g., suppository waxes), paraffins, silicones,talc, silicylate, etc. Each pharmaceutically acceptable diluent orcarrier used in a pharmaceutical composition of the invention must be“acceptable” in the sense of being compatible with the other ingredientsof the formulation and not injurious to the subject. Diluents orcarriers suitable for a selected dosage form and intended route ofadministration are well known in the art, and acceptable diluents orcarriers for a chosen dosage form and method of administration can bedetermined using ordinary skill in the art.

The pharmaceutical compositions of the invention may, optionally,contain additional ingredients and/or materials commonly used inpharmaceutical compositions. These ingredients and materials are wellknown in the art and include (1) fillers or extenders, such as starches,lactose, sucrose, glucose, mannitol, and silicic acid; (2) binders, suchas carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone,hydroxypropylmethyl cellulose, sucrose and acacia; (3) humectants, suchas glycerol; (4) disintegrating agents, such as agar-agar, calciumcarbonate, potato or tapioca starch, alginic acid, certain silicates,sodium starch glycolate, cross-linked sodium carboxymethyl cellulose andsodium carbonate; (5) solution retarding agents, such as paraffin; (6)absorption accelerators, such as quaternary ammonium compounds; (7)wetting agents, such as cetyl alcohol and glycerol monostearate; (8)absorbents, such as kaolin and bentonite clay; (9) lubricants, such astalc, calcium stearate, magnesium stearate, solid polyethylene glycols,and sodium lauryl sulfate; (10) suspending agents, such as ethoxylatedisostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters,microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agarand tragacanth; (11) buffering agents; (12) excipients, such as lactose,milk sugars, polyethylene glycols, animal and vegetable fats, oils,waxes, paraffins, cocoa butter, starches, tragacanth, cellulosederivatives, polyethylene glycol, silicones, bentonites, silicic acid,talc, salicylate, zinc oxide, aluminum hydroxide, calcium silicates, andpolyamide powder; (13) inert diluents, such as water or other solvents;(14) preservatives; (15) surface-active agents; (16) dispersing agents;(17) control-release or absorption-delaying agents, such ashydroxypropylmethyl cellulose, other polymer matrices, biodegradablepolymers, liposomes, microspheres, aluminum monostearate, gelatin, andwaxes; (18) opacifying agents; (19) adjuvants; (20) wetting agents; (21)emulsifying and suspending agents; (22), solubilizing agents andemulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate,ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol,1,3-butylene glycol, oils (in particular, cottonseed, groundnut, corn,germ, olive, castor and sesame oils), glycerol, tetrahydrofuryl alcohol,polyethylene glycols and fatty acid esters of sorbitan; (23)propellants, such as chlorofluorohydrocarbons and volatile unsubstitutedhydrocarbons, such as butane and propane; (24) antioxidants; (25) agentswhich render the formulation isotonic with the blood of the intendedrecipient, such as sugars and sodium chloride; (26) thickening agents;(27) coating materials, such as lecithin; and (28) sweetening,flavoring, coloring, perfuming and preservative agents. Each suchingredient or material must be “acceptable” in the sense of beingcompatible with the other ingredients of the formulation and notinjurious to the subject. Ingredients and materials suitable for aselected dosage form and intended route of administration are well knownin the art, and acceptable ingredients and materials for a chosen dosageform and method of administration may be determined using ordinary skillin the art.

The pharmaceutical compositions of the present invention suitable fororal administration may be in the form of capsules, cachets, pills,tablets, powders, granules, a solution or a suspension in an aqueous ornon-aqueous liquid, an oil-in-water or water-in-oil liquid emulsion, anelixir or syrup, a pastille, a bolus, an electuary or a paste. Theseformulations may be prepared by methods known in the art, e.g., by meansof conventional pan-coating, mixing, granulation or lyophilizationprocesses.

Solid dosage forms for oral administration (capsules, tablets, pills,dragees, powders, granules and the like) may be prepared, e.g., bymixing the active ingredient(s) with one or morepharmaceutically-acceptable diluents or carriers and, optionally, one ormore fillers, extenders, binders, humectants, disintegrating agents,solution retarding agents, absorption accelerators, wetting agents,absorbents, lubricants, and/or coloring agents. Solid compositions of asimilar type may be employed as fillers in soft and hard-filled gelatincapsules using a suitable excipient. A tablet may be made by compressionor molding, optionally with one or more accessory ingredients.Compressed tablets may be prepared using a suitable binder, lubricant,inert diluent, preservative, disintegrant, surface-active or dispersingagent. Molded tablets may be made by molding in a suitable machine. Thetablets, and other solid dosage forms, such as dragees, capsules, pillsand granules, may optionally be scored or prepared with coatings andshells, such as enteric coatings and other coatings well known in thepharmaceutical-formulating art. They may also be formulated so as toprovide slow or controlled release of the active ingredient therein.They may be sterilized by, for example, filtration through abacteria-retaining filter. These compositions may also optionallycontain opacifying agents and may be of a composition such that theyrelease the active ingredient only, or preferentially, in a certainportion of the gastrointestinal tract, optionally, in a delayed manner.The active ingredient can also be in microencapsulated form.

Liquid dosage forms for oral administration includepharmaceutically-acceptable emulsions, microemulsions, solutions,suspensions, syrups and elixirs. The liquid dosage forms may containsuitable inert diluents commonly used in the art. Besides inertdiluents, the oral compositions may also include adjuvants, such aswetting agents, emulsifying and suspending agents, sweetening,flavoring, coloring, perfuming and preservative agents. Suspensions maycontain suspending agents.

The pharmaceutical compositions of the present invention for rectal orvaginal administration may be presented as a suppository, which may beprepared by mixing one or more active ingredient(s) with one or moresuitable nonirritating diluents or carriers which are solid at roomtemperature, but liquid at body temperature and, therefore, will melt inthe rectum or vaginal cavity and release the active compound. Thepharmaceutical compositions of the present invention which are suitablefor vaginal administration also include pessaries, tampons, creams,gels, pastes, foams or spray formulations containing suchpharmaceutically-acceptable diluents or carriers as are known in the artto be appropriate.

Dosage forms for the topical or transdermal administration includepowders, sprays, ointments, pastes, creams, lotions, gels, solutions,patches, drops and inhalants. The active agent(s)/compound(s), includingthe malonate salt forms of the present invention, may be mixed understerile conditions with a suitable pharmaceutically-acceptable diluentor carrier. The ointments, pastes, creams and gels may containexcipients. Powders and sprays may contain excipients and propellants.

The pharmaceutical compositions of the present invention suitable forparenteral administrations may comprise one or more agent(s)/compound(s)in combination with one or more pharmaceutically-acceptable sterileisotonic aqueous or non-aqueous solutions, dispersions, suspensions oremulsions, or sterile powders which may be reconstituted into sterileinjectable solutions or dispersions just prior to use, which may containsuitable antioxidants, buffers, solutes which render the formulationisotonic with the blood of the intended recipient, or suspending orthickening agents. Proper fluidity can be maintained, for example, bythe use of coating materials, by the maintenance of the requiredparticle size in the case of dispersions, and by the use of surfactants.These pharmaceutical compositions may also contain suitable adjuvants,such as wetting agents, emulsifying agents and dispersing agents. It mayalso be desirable to include isotonic agents. In addition, prolongedabsorption of the injectable pharmaceutical form may be brought about bythe inclusion of agents which delay absorption.

In some cases, in order to prolong the effect of a drug (e.g.,pharmaceutical formulation), it is desirable to slow its absorption fromsubcutaneous or intramuscular injection. This may be accomplished by theuse of a liquid suspension of crystalline or amorphous material havingpoor water solubility.

The rate of absorption of the active agent/drug, including the malonatesalt forms of the present invention, then depends upon its rate ofdissolution which, in turn, may depend upon crystal size and crystallineform. Alternatively, delayed absorption of a parenterally-administeredagent/drug may be accomplished by dissolving or suspending the activeagent/drug in an oil vehicle. Injectable depot forms may be made byforming microencapsule matrices of the active ingredient inbiodegradable polymers. Depending on the ratio of the active ingredientto polymer, and the nature of the particular polymer employed, the rateof active ingredient release can be controlled. Depot injectableformulations are also prepared by entrapping the drug in liposomes ormicroemulsions which are compatible with body tissue. The injectablematerials can be sterilized for example, by filtration through abacterial-retaining filter.

The formulations may be present in unit-dose or multi-dose sealedcontainers, for example, ampules and vials, and may be stored in alyophilized condition requiring only the addition of the sterile liquiddiluent or carrier, for example water for injection, immediately priorto use. Extemporaneous injection solutions and suspensions may beprepared from sterile powders, granules and tablets of the typedescribed above.

The following examples are provided to further illustrate the compounds,compositions and methods of the present invention. These examples areillustrative only and are not intended to limit the scope of theinvention in any way.

EXAMPLES Example 1

Experimental Methods

X-Ray Powder Diffraction (XRPD)

Transmission mode XRPD patterns were collected using an incident beam ofCu radiation produced using a fine-focus source. An elliptically gradedmultilayer mirror was used to focus Cu Kα X-ray radiation through thespecimen and onto the detector. Prior to the analysis, a siliconspecimen (NIST SRM 640d) was analyzed to verify that the observedposition of the Si 111 peak was consistent with the NIST-certifiedposition. A specimen of the sample was sandwiched between 3-μm-thickfilms and analyzed in transmission geometry. A beam-stop, shortantiscatter extension, and antiscatter knife edge were used to minimizethe background generated by air. Soller slits for the incident anddiffracted beams were used to minimize broadening from axial divergence.Diffraction patterns were collected using a scanning position-sensitivedetector located 240 mm from the specimen. Preferred orientation andparticle static effects were not assessed.

Reflection mode XRPD patterns were collected using an incident beam ofCu Kα radiation produced using a fine-focus source and a nickel filter.The diffractometer was configured using the symmetric Bragg-Brentanogeometry. Prior to the analysis, a silicon specimen (NIST SRM 640d) wasanalyzed to verify that the observed position of the Si 111 peak wasconsistent with the NIST-certified position. A specimen of the samplewas prepared as a thin, circular layer centered on a siliconzero-background substrate. Antiscatter slits (SS) were used to minimizethe background generated by air. Soller slits for the incident anddiffracted beams were used to minimize broadening from axial divergence.Diffraction patterns were collected using a scanning position-sensitivedetector located 240 mm from the sample. Preferred orientation andparticle static effects were not assessed.

Under most circumstances, peaks within the range of up to about 30° 2θwere selected. The location of the peaks along the x-axis (° 2θ) wererounded to one significant figure after the decimal point. Peak positionvariabilities are given to within ±0.2° 2θ based upon recommendationsoutlined in the USP discussion of variability in X-ray powderdiffraction. The accuracy and precision associated with any particularmeasurement was not determined. Moreover, third party measurements onindependently prepared samples on different instruments may lead tovariability which is greater than ±0.2° 2θ. Per USP guidelines, variablehydrates and solvates may display peak variances greater than 0.2° 2θand therefore peak variances of 0.2° 2θ are not applicable to thesematerials. For d-space listings, the wavelength used to calculated-spacings was 1.5405929 Å, the Cu—Kα1 wavelength. Variabilityassociated with d-spacing estimates was calculated from the USPrecommendation, at each d-spacing, and provided in the respective datatables.

Fourier Transform Infrared (FT-IR) Spectroscopy

FT-IR spectra were acquired using a Fourier transform infraredspectrophotometer equipped with a mid/far IR source, an extended rangepotassium bromide (KBr) beamsplitter, and a deuterated triglycinesulfate (DTGS) detector. Wavelength verification was performed usingNIST SRM 1921b (polystyrene). An attenuated total reflectance (ATR)accessory with a germanium (Ge) crystal was used for data acquisition.256 co-added scans were collected at a spectral resolution of 2 cm⁻¹. Abackground data set was acquired with a clean Ge crystal. A Log 1/R(R=reflectance) spectrum was obtained by taking a ratio of these twodata sets against each other. Peak picking was performed using anabsolute threshold near the baseline and a sensitivity of 75.

Differential Scanning Calorimetry (DSC)

DSC analysis was performed using a differential scanning calorimeter.Temperature calibration was performed using NIST-traceable indium metal.The sample was placed into an aluminum DSC pan, covered with a lid, andthe weight was accurately recorded. A weighed aluminum T0HSMP panconfigured as the sample pan was placed on the reference side of thecell. Reported temperatures are rounded to 1 degree unless specifiedotherwise.

Example 2 Preparation of Crystalline Free Base4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide

4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide free base was preparedaccording to the following synthesis scheme.

In Step 1, a clean and dry 200 L glass-lined reactor was evacuated to≦−0.08 MPa, and then filled with nitrogen to normal pressure threetimes. Anhydrous ethanol (49.90 kg) was charged into the 200 Lglass-lined reactor. ASYM-111606 (Asymchem) (12.70 kg) andisopropylamine (29.00 kg) were added into the mixture in turn. Themixture was heated to 65-75° C. for refluxing. The mixture reacted at65-75° C. After 20 h, the reaction was sampled and analyzed by HPLCevery 4-6 h until the content of ASYM-111606 was ≦1%. The mixture wascooled to 40-45° C. and was concentrated at ≦45° C. under reducedpressure (≦−0.08 MPa) until 13-26 L remained. The organic phase waswashed with a sodium chloride solution and was stirred for 20-30 min andsettled for 20-30 min before separation. The organic phase wasconcentrated at ≦30° C. under reduced pressure (≦−0.06 MPa) until 13-20L remained. Petroleum ether (8.55 kg) was added into the concentratedmixture. The mixture was transferred into a 20 L rotary evaporator andcontinued concentrating at ≦30° C. under reduced pressure (≦−0.06 MPa)until 13-20 L remained. Then petroleum ether (8.55 kg) was added intothe concentrated mixture. The mixture was cooled to 0-5° C. and stirredfor crystallization. After 1 h, the mixture was sampled and analyzed bywt % every 1-2 h until the wt % of the mother liquor was ≦11%, or thechange of the wt % between consecutive samples was ≦1%. The mixture wasfiltered with a 10 L filter flask. The filter cake was sampled andanalyzed for purity by HPLC. 10.50 kg of product was recovered as abrownish yellow solid at 99.39% purity.

In Step 2, a clean and dry 300 L glass-lined reactor was evacuated to≦−0.08 MPa, and then filled with nitrogen to normal pressure threetimes. Glycol dimethyl ether (73.10 kg) was charged into the 300 Lglass-lined reactor at 20-30° C. ASYM-112060 (Asymchem) (10.46 kg) andASYM-111938 (Asymchem) (12.34 kg, 11.64 kg after corrected) were addedinto the mixture in turn under the protection of nitrogen. Maintainingthe temperature at 20-30° C., purified water (10.50 kg) and anhydroussodium carbonate (5.67 kg) were added into the mixture. Palladiumacetate (0.239 kg) and tricyclohexylphosphonium tetrafluoroborate (0.522kg) were added into the mixture under the protection of nitrogen. Afteraddition, the mixture was evacuated to ≦−0.06 MPa, and then filled withnitrogen to normal pressure. This was repeated for ten times untilresidual oxygen was ≦300 ppm. The mixture was heated to 75-85° C. forrefluxing. The mixture reacted at 75-85° C. After 4 h, the mixture wassampled and analyzed by HPLC every 2-3 h for content of ASYM-112060. Thecontent of ASYM-112060 was 6.18%, so additional ASYM-111938 (0.72 kg)was added and continued reaction until the content of ASYM-112060 was≦3%. The mixture was cooled to 25-35° C. and filtered with a 30 Lstainless steel vacuum filter. The filter cake was soaked and washedtwice with THF (14.10 kg). The filtrate and washing liquor were combinedand concentrated at ≦50° C. under reduced pressure (≦−0.08 MPa) until10-15 L remained. The mixture was cooled to 15-25° C. Methanol (11.05kg) was added into the concentrated mixture. Then the mixture wasstirred for crystallization. After 2 h, the mixture was sampled andanalyzed by HPLC every 2-4 h until the wt % of the mother liquor was≦2%. The mixture was filtered with a 30 L stainless steel vacuum filter.The filter cake was soaked and washed twice with methanol (8.30 kg). Thefilter cake was transferred into a 50 L plastic drum. Then ethyl acetate(7.10 kg) and petroleum ether (46.30 kg) were added into the drum. Themixture was stirred for 1.5-2 h and then filtered with a nutsche filter.The filter cake was soaked and washed with petroleum ether (20.50 kg).The filter cake was dried in the nutsche filter under nitrogen at 30-40°C. After 8 h, the solid was sampled and Karl Fischer (KF) analysis wasperformed in intervals of 4-8 h to monitor the drying process. Dryingwas completed when the KF result was ≦1.0% water. During drying, thesolid was turned over and mixed every 4-6 h. 12.15 kg of product wasrecovered as a brownish yellow solid at 98.32% purity.

In Step 3, a clean and dry 300 L glass-lined reactor was evacuated to≦−0.08 MPa, and then filled with nitrogen to normal pressure threetimes. THF (62.58 kg) was charged into the 300 L glass-lined reactor at15-30° C. Then the stirrer was started. ASYM-112393 (12.00 kg, 11.70 kgafter corrected) was added into the mixture. The mixture was stirreduntil the solid dissolved completely. Maintaining the temperature at15-30° C., a lithium hydroxide solution which was prepared with lithiumhydroxide monohydrate (5.50 kg) in purified water (70.28 kg) was addedinto the mixture. Then diethylamine (3.86 kg) was added. The mixture washeated to 60-70° C. for refluxing. The mixture reacted at 60-70° C.After 30 h, the reaction was sampled and analyzed by HPLC every 4-6 huntil the content of intermediate at relative retention time(RRT)=1.39-1.44 was <1% and the content of ASYM-112393 was <1%. HPLCconditions for this analysis are set forth in Table 1.

TABLE 1 HPLC Parameters Column: ACE 3 C18, 4.6 × 150 mm, (ACE-111-1546)Column 30° C. Temperature: Flow rate 1.1 mL/min Injection 10 μL Volume:Mobile Phase A: 0.05% TFA in water (v/v) Mobile Phase B: 0.05% TFA inAcetonitrile (v/v) Gradient Table: T(min): B% 0.0 5 4.0 20 14.0 85 14.15 18.5 5 Detection: UV at 215 nm Run time 18.5 min

The mixture was cooled to 25-35° C. and MTBE (25.97 kg) was added intothe mixture. The mixture was stirred for 20-30 min and filtered via anin-line fluid filter. The filtrate was transferred into a 300 Lglass-lined reactor and settled for 20-30 min before separation. The pHof the obtained aqueous phase was adjusted with a 6 N hydrochloric acidsolution which was prepared from concentrated hydrochloric acid (14.86kg) in purified water (10.88 kg) at the rate of 5-8 kg/h at 15-25° C.until the pH was 1-2. The pH of the mixture was adjusted again with asaturated sodium carbonate solution which was prepared from sodiumcarbonate (5.03 kg) in purified water (23.56 kg) at the rate of 3-5 kg/hat 15-25° C. until the pH was 6.4-6.7. Then the pH of the mixture wasadjusted with a hydrochloric acid solution which was prepared fromconcentrated hydrochloric acid (1.09 kg) in purified water (0.80 kg)until the pH was 6.2-6.4. The mixture was filtered with a nutschefilter. The filter cake was transferred into a 300 L glass-lined reactorand purified water (117.00 kg) was added. The mixture was stirred andsampled and analyzed by HPLC until the p-toluenesulfonic acid residue ofthe filter cake was ≦0.5%. Then the mixture was filtered. The filtercake was dried in the tray drier under nitrogen at 55-65° C. untilKF≦10%. The solid and MTBE (8.81 kg) were charged into a 50 L stainlesssteel drum. The mixture was stirred for 1-2 h. The mixture was filteredwith a 30 L stainless steel vacuum filter. The filter cake was dried inthe nutsche filter at 50-60° C. After 8 h, the solid was sampled andanalyzed by KF every 4-8 h until KF≦5%. During drying, the solid wasturned over and mixed every 4-6 h. 6.3 kg of product was recovered as anoff-white solid at 98.07% purity.

In Step 4, a dry and clean 50 L flask was purged with nitrogen for 20min. DMF (30.20 kg) was charged into the 50 L flask reactor. Then thestirrer was started. Maintaining the temperature at 15-25° C.,ASYM-112394 (3.22 kg, 2.76 kg after corrected) was added into themixture. The mixture was stirred until the solid dissolved completely.The mixture was cooled to −10 to −20° C. and 1-hydroxybenzotriazolehydrate (2.10 kg) was added into the mixture at −10 to −20° C. Then EDCl(2.41 kg) was added into the mixture in five portions at an interval ofabout 5-10 min. The mixture was cooled to −20 to −30° C. and ASYM-111888(Asymchem) (1.96 kg) was added into the mixture at −20 to −30° C. ThenDIEA (1.77 kg) was added into the mixture at the rate of 3-4 kg/h. Themixture was heated to 15-25° C. at the rate of 5-10° C./h. The mixturewas reacted at 15-25° C. After 6-8 h, the mixture was sampled andanalyzed by HPLC every 2-4 h until the content of ASYM-112394 was ≦2%.The mixture was cooled to 0-10° C. and the reaction mixture was quenchedwith a solution which was prepared from ethyl acetate (28.80 kg) inpurified water (12.80 kg) at 0-10° C. The mixture was extracted threetimes with ethyl acetate (28.80 kg). For each extraction the mixture wasstirred for 20-30 min and settled for 20-30 min before separation. Theorganic phases were combined and washed twice with purified water (12.80kg). The mixture was stirred for 20-30 min and settled for 20-30 minbefore separation for each time. Then the obtained organic phase wasfiltered through an in-line fluid filter. The filtrate was transferredinto a 300 L glass-lined reactor. The mixture was washed twice with a 5%acetic acid solution, which was prepared from acetic acid (2.24 kg) inpurified water (42.50 kg). The solution was added at the rate of 10-20kg/h. The organic phase was washed twice with a sodium carbonatesolution, which was prepared from sodium carbonate (9.41 kg) in purifiedwater (48.00 kg). The organic phase was washed twice with a sodiumchloride solution, which was prepared from sodium chloride (16.00 kg) inpurified water (44.80 kg). The organic phase was transferred into a 300L glass-lined reactor. Anhydrous sodium sulfate (9.70 kg) was added intothe mixture and the mixture was stirred for 2-4 h at 15-30° C. Themixture was filtered with a nutsche filter, which was pre-loaded withabout 1 cm thick silica gel (7.50 kg). The filter cake was soaked andwashed with ethyl acetate (14.40 kg) before filtration. The filtrateswere combined and the combined filtrate was added into a 72 L flaskthrough an in-line fluid filter. The mixture was concentrated at T≦40°C. under reduced pressure (P≦−0.08 MPa) until 3-4 L remained. Then MTBE(4.78 kg) was added into the mixture. The mixture was cooled to 0-10° C.for crystallization with stirring. After 1 h, the mixture was sampledand analyzed by wt % every 1-2 h until the wt % of the mother liquor was≦5%, or the change of wt % between consecutive samples was ≦1%. Themixture was filtered with a vacuum filter flask and the filter cake wasdried in the tray drier under nitrogen at 30-40° C. until KF≦0.5%. 3.55kg of product was recovered as an off-white solid at 100% purity.

The resulting4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide free base was analyzed by XRPD(FIG. 1). Peaks shown in FIG. 1 are listed in Table 2, prominent peaksare listed in Table 3.

TABLE 2 XRPD peaks observed for 4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide free base. 2θ (°) d space (Å)Intensity (%)  9.1 ± 0.2 9.690 ± 0.212 12 10.0 ± 0.2 8.869 ± 0.178 210.2 ± 0.2 8.664 ± 0.169 7 11.4 ± 0.2 7.742 ± 0.135 5 12.5 ± 0.2 7.066 ±0.112 25 12.7 ± 0.2 6.956 ± 0.109 8 13.3 ± 0.2 6.637 ± 0.099 2 15.2 ±0.2 5.833 ± 0.076 15 15.4 ± 0.2 5.769 ± 0.075 46 16.0 ± 0.2 5.531 ±0.069 9 17.1 ± 0.2 5.173 ± 0.060 3 17.6 ± 0.2 5.038 ± 0.057 8 18.2 ± 0.24.876 ± 0.053 4 18.8 ± 0.2 4.723 ± 0.050 2 19.2 ± 0.2 4.624 ± 0.048 1219.5 ± 0.2 4.556 ± 0.046 100 20.3 ± 0.2 4.381 ± 0.043 14 20.5 ± 0.24.327 ± 0.042 12 21.4 ± 0.2 4.145 ± 0.038 44 21.7 ± 0.2 4.102 ± 0.037 1121.9 ± 0.2 4.057 ± 0.037 12 23.1 ± 0.2 3.847 ± 0.033 13 23.3 ± 0.2 3.812± 0.032 25 23.6 ± 0.2 3.774 ± 0.032 26 24.3 ± 0.2 3.653 ± 0.030 11 25.2± 0.2 3.530 ± 0.028 9 25.6 ± 0.2 3.476 ± 0.027 2 26.6 ± 0.2 3.355 ±0.025 3 27.0 ± 0.2 3.297 ± 0.024 7 27.7 ± 0.2 3.214 ± 0.023 13 27.9 ±0.2 3.191 ± 0.022 10 28.2 ± 0.2 3.159 ± 0.022 3 28.7 ± 0.2 3.106 ± 0.0219 28.9 ± 0.2 3.083 ± 0.021 4 29.2 ± 0.2 3.057 ± 0.020 9 30.2 ± 0.2 2.957± 0.019 14 30.6 ± 0.2 2.923 ± 0.019 9

TABLE 3 Prominent XRPD peaks for4-(5-Chloro-2-isopropylaminopyridin-4-yl)- 1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl] amide free base. 2θ (°) d space (Å)Intensity (%)  9.1 ± 0.2 9.690 ± 0.212 12 12.5 ± 0.2 7.066 ± 0.112 2515.2 ± 0.2 5.833 ± 0.076 15 15.4 ± 0.2 5.769 ± 0.075 46 19.2 ± 0.2 4.624± 0.048 12 19.5 ± 0.2 4.556 ± 0.046 100 20.3 ± 0.2 4.381 ± 0.043 14 20.5± 0.2 4.327 ± 0.042 12 21.4 ± 0.2 4.145 ± 0.038 44 21.7 ± 0.2 4.102 ±0.037 11 21.9 ± 0.2 4.057 ± 0.037 12 23.1 ± 0.2 3.847 ± 0.033 13 23.3 ±0.2 3.812 ± 0.032 25 23.6 ± 0.2 3.774 ± 0.032 26 24.3 ± 0.2 3.653 ±0.030 11 27.7 ± 0.2 3.214 ± 0.023 13 27.9 ± 0.2 3.191 ± 0.022 10 30.2 ±0.2 2.957 ± 0.019 14

FT-IR was performed on a sample of4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide free base as described inExample 1 (FIG. 2). Observed peaks from FIG. 2 are listed in Table 4.

TABLE 4 Observed FT-IR peaks for4-(5-Chloro-2-isopropylaminopyridin-4-yl)- 1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl] amide free base. Position (cm⁻¹)Intensity 681 0.0174 712 0.0025 748 0.0014 783 0.0058 807 0.001 8270.0082 857 0.0045 878 0.00069 897 0.00067 916 0.00056 932 0.0008 9960.0004 1040 0.00074 1080 0.0069 1101 0.00081 1126 0.00096 1145 0.00141170 0.0027 1197 0.0011 1208 0.0028 1235 0.0013 1255 0.0015 1268 0.00211294 0.0013 1350 0.0018 1364 0.002 1385 0.00077 1398 0.00077 1439 0.00171451 0.0014 1466 0.0019 1487 0.0089 1504 0.0033 1523 0.0065 1533 0.00631568 0.0021 1603 0.0108 1629 0.0062 2927 0.00024 2974 0.00028 32350.00052 3405 0.00026

DSC was performed on a sample of4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide free base as described inExample 1 (FIG. 3) and showed an endotherm having an onset temperatureof approximately 184° C.

Example 3A Preparation of4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide Form C

4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide Form C was prepared from4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide free base as follows.ASYM-111935 (10.4 kg) was added to a stirred mixture of anhydrousethanol (73.9 kg), methanol (4.1 kg) and isopropanol (4.1 kg). Themixture was heated to 70-75° C. and stirred until all the solidsdissolved. Anhydrous HCl (37 wt %, 1.1 eq) in a mixture ofethanol/methanol/isopropanol (90:5:5) was added and the mixturemaintained at 70-75° C. for 2 hours after the addition was completed.The mixture was then cooled to 15-25° C. at a rate of 5-15° C. per hourand stirred at this temperature until the desired polymorphic purity wasreached. The end point of the crystallization/polymorph conversion wasdetermined by the absence of an XRPD peak at about 10.5° 2θ in threesuccessive samples.

The mixture was then filtered and washed successively with apre-prepared solution of anhydrous ethanol (14.8 kg), methanol (0.8 kg)and isopropanol (0.8 kg), followed by MTBE (2×21 kg). Avoidance of delayin the washing of the filter cake is preferable because the polymorphmay be unstable in the wet filter cake in the presence of reagentalcohol and improved stability was observed after the MTBE wash has beenperformed. The wet filter cake was then dried in a heated filter funnelor a tray drier at 40-50° C. until dry. Typical yields were about85-90%.

Example 3B Alternative Preparation of4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide Form C

4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide Form C was also prepared from4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide free base as follows. A dry andclean 72 L flask was purged with nitrogen for 20 min. Anhydrous ethanol(21.35 kg) methanol (1.17 kg) and isopropanol (1.19 kg) were chargedinto the 72 L flask at 15-25° C. and the mixture was stirred for 20-30min. ASYM-111935 (3.01 kg) was added into the mixture and heated to70-75° C. at the rate of 15-25° C./h and stirred until the soliddissolved completely.

An alcohol/HCl solution was prepared as follows. Anhydrous ethanol(1.500 kg) methanol (0.088 kg) and isopropanol (0.087 kg) were chargedinto a 5 L flask at 15-25° C. and the mixture was stirred for 20-30 min.The mixture was bubbled with hydrogen chloride through a dip tube understirring at 10-25° C. After 2 h, the mixture was sampled and analyzedevery 2-4 h until the wt % of hydrogen chloride was ≧35%.

The alcohol/HCl solution (0.519 kg) prepared above was added dropwiseinto the mixture at the rate of 0.5-1.0 kg/h at 70-75° C. Seed crystal(0.009 kg) was added into the mixture and the alcohol/HCl solution(0.173 kg) prepared above was added into the mixture at the rate of0.5-1.0 kg/h at 70-75° C. After addition, the mixture was stirred for1-2 h at 70-75° C. The mixture was cooled to 15-25° C. at the rate of5-15° C./h and stirred for 4-6 h. The mixture was heated to 70-75° C. atthe rate of 15-25° C./h and stirred for 8-10 h at 70-75° C. The mixturewas cooled to 15-25° C. at the rate of 5-15° C./h and stirred for 4-6 h.The mixture was filtered with a vacuum filter flask. The filter cake wassoaked and rinsed with a solution which was prepared from anhydrousethanol (4.25 kg) and methanol (0.24 kg) and isopropanol (0.24 kg)before filtration. The filter cake was dried in a drying room undernitrogen at 40-50° C. until the ethanol residue was <0.5% and methanolresidue was <0.3% and isopropanol residue was <0.3%. 2.89 kg of productwas recovered as a white solid at 99.97% purity.

The resulting4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide Form C was analyzed by XRPD(FIG. 4). Peaks shown in FIG. 4 are listed in Table 5, prominent peaksare listed in Table 6.

TABLE 5 XRPD peaks observed for4-(5-Chloro-2-isopropylaminopyridin-4-yl)- 1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl] amide Form C. 2θ (°) d space (Å)Intensity (%)  6.1 ± 0.2 14.436 ± 0.472  17  6.7 ± 0.2 13.099 ± 0.388 61  8.6 ± 0.2 10.287 ± 0.239  5 10.8 ± 0.2 8.196 ± 0.152 5 11.0 ± 0.28.039 ± 0.146 15 12.1 ± 0.2 7.335 ± 0.121 15 12.4 ± 0.2 7.108 ± 0.114 613.5 ± 0.2 6.533 ± 0.096 8 13.7 ± 0.2 6.467 ± 0.094 10 15.2 ± 0.2 5.828± 0.076 38 16.5 ± 0.2 5.363 ± 0.064 18 16.9 ± 0.2 5.258 ± 0.062 7 17.2 ±0.2 5.139 ± 0.059 5 17.6 ± 0.2 5.023 ± 0.056 59 17.9 ± 0.2 4.949 ± 0.05537 18.4 ± 0.2 4.818 ± 0.052 32 18.7 ± 0.2 4.743 ± 0.050 13 19.0 ± 0.24.671 ± 0.049 4 19.2 ± 0.2 4.628 ± 0.048 4 19.6 ± 0.2 4.529 ± 0.046 1419.9 ± 0.2 4.450 ± 0.044 100 20.4 ± 0.2 4.354 ± 0.042 18 20.6 ± 0.24.318 ± 0.042 28 20.8 ± 0.2 4.272 ± 0.041 52 21.5 ± 0.2 4.122 ± 0.038 2822.1 ± 0.2 4.016 ± 0.036 4 22.6 ± 0.2 3.935 ± 0.034 28 22.7 ± 0.2 3.923± 0.034 27 23.5 ± 0.2 3.785 ± 0.032 43 24.0 ± 0.2 3.704 ± 0.030 29 24.3± 0.2 3.664 ± 0.030 12 24.5 ± 0.2 3.634 ± 0.029 8 24.9 ± 0.2 3.573 ±0.028 56 25.4 ± 0.2 3.498 ± 0.027 60 25.7 ± 0.2 3.467 ± 0.027 37 26.0 ±0.2 3.424 ± 0.026 6 26.4 ± 0.2 3.375 ± 0.025 8 27.7 ± 0.2 3.224 ± 0.02322 28.0 ± 0.2 3.182 ± 0.022 11 28.3 ± 0.2 3.147 ± 0.022 8 29.2 ± 0.23.056 ± 0.020 4 29.6 ± 0.2 3.020 ± 0.020 7 29.9 ± 0.2 2.983 ± 0.019 2830.2 ± 0.2 2.957 ± 0.019 10

TABLE 6 Prominent XRPD peaks for4-(5-Chloro-2-isopropylaminopyridin-4-yl)- 1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl] amide Form C. 2θ (°) d space (Å)Intensity (%)  6.1 ± 0.2 14.436 ± 0.472  17  6.7 ± 0.2 13.099 ± 0.388 61 11.0 ± 0.2 8.039 ± 0.146 15 12.1 ± 0.2 7.335 ± 0.121 15 13.7 ± 0.26.467 ± 0.094 10 15.2 ± 0.2 5.828 ± 0.076 38 16.5 ± 0.2 5.363 ± 0.064 1817.6 ± 0.2 5.023 ± 0.056 59 17.9 ± 0.2 4.949 ± 0.055 37 18.4 ± 0.2 4.818± 0.052 32 18.7 ± 0.2 4.743 ± 0.050 13 19.6 ± 0.2 4.529 ± 0.046 14 19.9± 0.2 4.450 ± 0.044 100 20.4 ± 0.2 4.354 ± 0.042 18 20.6 ± 0.2 4.318 ±0.042 28 20.8 ± 0.2 4.272 ± 0.041 52 21.5 ± 0.2 4.122 ± 0.038 28 22.6 ±0.2 3.935 ± 0.034 28 22.7 ± 0.2 3.923 ± 0.034 27 23.5 ± 0.2 3.785 ±0.032 43 24.0 ± 0.2 3.704 ± 0.030 29 24.3 ± 0.2 3.664 ± 0.030 12 24.9 ±0.2 3.573 ± 0.028 56 25.4 ± 0.2 3.498 ± 0.027 60 25.7 ± 0.2 3.467 ±0.027 37 27.7 ± 0.2 3.224 ± 0.023 22 28.0 ± 0.2 3.182 ± 0.022 11 29.9 ±0.2 2.983 ± 0.019 28 30.2 ± 0.2 2.957 ± 0.019 10

FT-IR was performed on a sample of Form C as described in Example 1(FIG. 5). Observed peaks from FIG. 5 are listed in Table 7.

TABLE 7 Observed FT-IR peaks for4-(5-Chloro-2-isopropylaminopyridin-4-yl)- 1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl] amide Form C. Position (cm⁻¹)Intensity 680 0.0389 694 0.0737 705 0.0203 723 0.0273 728 0.0245 7420.0263 771 0.0449 785 0.0527 845 0.0479 865 0.0128 879 0.0232 922 0.0112946 0.0275 958 0.011 985 0.0119 1000 0.0124 1076 0.0649 1107 0.0183 11290.0245 1141 0.0322 1177 0.018 1219 0.0554 1246 0.0238 1282 0.0279 13100.0342 1324 0.0179 1344 0.0144 1376 0.0239 1380 0.024 1389 0.0204 14130.0196 1436 0.0324 1472 0.0279 1498 0.0254 1523 0.0543 1551 0.027 15740.0371 1610 0.0697 1643 0.0865 2952 0.0153 2977 0.0167 3057 0.015 31780.0147 3229 0.0162 3294 0.0171 3369 0.0161

DSC was performed on a sample of Form C as described in Example 1 (FIG.6) and showed a prominent endotherm having onset temperature ofapproximately 239° C.

Example 44-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide Salt Screen

Salt formers for screening were selected based on the pKa predicted for4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide free base (about 5). Molarratios of the starting material to salt formers at mixing were about 1:1for the majority of the experiments. Selected experiments withappropriate acids were conducted with approximately two fold excess ofthe free base. Experiments were designed based on kinetic solubilitiesof starting materials estimated for this purpose by solvent additionmethod. Mostly cooling and slurry techniques or combinations thereofwere employed primarily at medium scale of starting material (˜50-100mg). XRPD was the primary analytical technique for initialidentification of new materials. XRPD patterns acquired on isolatedsolids were compared to each other, the pattern of the free base andavailable patterns of salt formers.

Table 8 summarizes the salt formers tested, conditions used, observedresults and a description of preliminary XRPD analyses. Table 9 definesabbreviations used in this series of experiments.

TABLE 84-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide salt screen. Salt former XRPD(ratio) Conditions Observations Results Adipic acid Mixed hot solutionof acid in EtOH with free Unknown Free base (1:1) base at 68° C. Slurry,RT, overnight. morphology, birefringence/ extinction L-Aspartic Mixedhot solution of free base in EtOH with Unknown Free base + acid hot aq.Acid at 65-70° C. (EtOH/water, 40/60). morphology, L-aspartic (1:1)Slurry, 70° C., 30 min then RT, 1 hour. birefringence/ acid extinctionBSA Mixed hot solution of free base in No solids N/A (1:1) EtOH/heptane,80/20 with acid at 60° C. Stirring, RT, 1 day. Sonicated, cooled to −15°C. to −25° C. overnight. VF attempt (gummy). Re-dissolved in MeOH, FE.Mixed solution of acid in EtOH with free base Unknown X-ray (mostlyclear, few solids remained). Brought morphology, amorphous to 65° C.(clear). Added MTBE until cloudy, birefringence/ (EtOH/MTBE 20/80),sonicated. Stirring, RT, extinction overnight (viscous). Brought to 60°C. (dissolved), evaporated to about ½ volume, SC to RT. Sonicated,cooled to −15° C. to −25° C., 4 days. Benzoic acid Mixed hot solution ofacid in EtOH at 68° C. Unknown Free base (1:1) with free base. Slurry,RT, overnight. morphology, birefringence/ extinction Citric acid EtOH,slurry attempt, RT. Cooled to −15° C. No solids N/A (1:1) to −25° C.overnight. Mixed hot suspension of free base in EtOAc Unknown Freebase + with acid at 65° C. Slurry, 65° C. then RT, morphology, citricacid overnight. birefringence/ extinction EDSA Mixed hot solution ofacid in MEK/heptane Unknown X-ray (dihydrate) 80/20 with free base at60° C. (clumped up morphology, amorphous (1:1) and became sticky, nochange after some sonication). Slurry, 67° C. then RT overnight.birefringence/ Decanted. IPA slurry, RT, 20 min. extinction EDSA Mixedhot solution of acid in MEK/heptane Unknown X-ray (dihydrate) 80/20 withfree base at 60° C. Slurry, 67° C. morphology, amorphous (2:1) then RT,overnight. some birefringence/ extinction ESA Mixed hot solution of freebase in ACN with No solids N/A (1:1) acid at 60° C. Stirring, RT, 1 day.Sonicated, cooled to −15° C. to −25° C., 6 days. Mixed hot yellowsolution of free base in IPA No solids N/A with acid at 73° C.(clear/colorless). Stirring, 75° C. then RT. Sonicated with partialevaporation, RT Mixed hot yellow solution of free base in IPA No solidsN/A with acid at 73° C. (clear/colorless). Stirring, 75° C. then RT.Sonicated with partial evaporation, RT. Added water, IPA/water 71/29.Cooled to 2-8° C., 10 days. Fumaric acid EtOH, slurry, RT. Unknown Freebase (1:1) morphology, birefringence/ extinction D-Gluconic Mixedsuspension of free base in EtOH with Unknown Free base acid (about 50%aq. acid. Stirring, RT, 2 days. Cooled to morphology, aqueous 2-8° C.,overnight. Added water (EtOH/water birefringence/ solution) 30/70).Slurry, RT, overnight. extinction (1:1) α Ketoglutaric EtOH, slurryattempt, RT. Cooled to −15° C. No solids N/A acid to −25° C., overnight.(1:1) Mixed hot solution of free base in IPA with Unknown X-ray acid at60° C. Stirring, RT, 1 hour. morphology, amorphous + Sonicated, cooledto 2-8° C., overnight. birefringence/ small extinction peaks L-Malicacid EtOH, slurry attempt, RT. No solids N/A (1:1) Malonic acid EtOH,slurry, RT, 1 day. VF attempt, Needles, Malonate (1:1) evaporation offiltrate. birefringence/ Form A extinction MSA Mixed hot solution offree base in ACN with Oil N/A (1:1) acid at 60° C. (gummy solids).Stirring, RT, 1 day, sonicated. Redissolved in MeOH), SE. Mixed hotsolution of free base in ACN with No solids N/A acid at 60° C. (gummysolids). Stirring, RT, 1 day, sonicated. Redissolved in MeOH), SE.Slurry attempt with IPA, sonicated. Cooled to −15° C. to −25° C.overnight. RT, 1 week. Mixed hot solution of free base (slightly Nosolids N/A cloudy, yellow) in IPA with acid at 73° C. (clear/colorless).Stirring, 75° C. then RT. Sonicated with partial evaporation, RT Mixedhot solution of free base (slightly No solids N/A cloudy, yellow) in IPAwith acid at 73° C. (clear/colorless). Stirring, 75° C. then RT.Sonicated with partial evaporation, RT. Added water, IPA/water 83/17Added acid to suspension of free base in No solids N/A EtOH/MTBE 30/70(gummy solids beneath clear/colorless solution). Stirring, RT, 1 day.Added acid to suspension of free base in Unknown X-ray ACN/MTBE 50/50(gummy solid beneath morphology, amorphous clear/colorless solution).Stirring, RT, 1 day. solvent present, some glassy particles, somebirefringence/ extinction Added acid to suspension of free base inUnknown X-ray MIBK (gummy solid beneath clear/colorless morphology,amorphous solution). Stirring, RT, 1 day. some glassy particles, somebirefringence/ extinction Mixed hot suspension of free base in No solidsN/A MCH/MeOH 90/10 with acid at 80° C. (some viscous). SC, RT, 1 day.Oxalic acid Mixed hot suspension of free base in EtOAc Unknown Free base(1:1) with acid at 65° C. Some solids remained. morphology, Stirring,65° C. then RT, 1 day (solution, birefringence/ solids on walls).extinction Mixed solution of acid in EtOH with free Unknown Unique,base. Slurry, RT, sonicated. Added MTBE morphology, crystalline(EtOH/MTBE 55/45). Stirring, RT, overnight. possible birefringence/extinction Phosphoric EtOH, slurry, RT. Unknown X-ray acid morphology,amorphous + (1:1) some crystalline birefringence/ free base extinctionMixed hot solution of free base in EtOH Unknown N/A (cloudy, yellow)with acid at at 73° C. (clear). morphology, Stirring, 75° C., 10 minthen RT (viscous possible residue). Added MTBE (solids clumped up),birefringence/ decanted. VO, RT, overnight. extinction + glassy Mixedhot solution of free base in EtOH Unknown Free base (cloudy, yellow)with acid at at 73° C. (clear). morphology, Stirring, 75° C., 10 minthen RT (viscous some residue). Added MTBE (solids clumped up),birefringence/ decanted. VO, RT, overnight. Slurry, EtOH/ extinctionwater 30/70, 75° C., 15 min (clear then some solids). Cooled to 65° C.,30 min (more solids present). Cooled to 55° C., 1 hr. (increase insolids). Cooled to 45° C., 1 hr. then RT overnight. Succinic acid EtOH,slurry, RT, 1 day. VF attempt, Unknown Free base (1:1) evaporation offiltrate. morphology, birefringence/ extinction Sulfuric acid Mixed hotsolution of free base in ACN with Unknown Unique, (1:1) acid at 60° C.(gummy). Stirring, 60° C. then morphology, poorly RT, 1 day. Sonicated,decanted. IPA slurry some crystalline with sonication, RT, overnight.birefringence/ extinction Mixed hot solution of free base in ACN withUnknown Same as acid at 60° C. (gummy). Stirring, 60° C. morphology,above, then RT, 1 day. Sonicated, decanted. IPA possible poorly slurrywith sonication, RT, overnight. Slurry, birefringence/ crystallineIPA/heptane 50/50, RT, 4 days. extinction Mixed hot suspension of freebase in IPA Viscous N/A with acid at 60° C. (mostly dissolved, somesubstance viscous). Cool to RT, slurry attempt (viscous). Attempt tore-dissolve in various organic solvents. Mixed hot suspension of freebase in EtOAc Viscous N/A with acid 65° C. (sticky solids). VF whilehot, 2 hr. substance Mixed hot suspension of free base in Oil, no solidsN/A nitromethane with acid at 65° C. (sticky solids, slightly pinksolution). Cool to RT, stirring, 3 hr. Mixed hot solution of free basein EtOH with Viscous residue N/A acid at 65-75° C. (clear). Cool to RT,stirring, 5 days. Mixed hot suspension of free base in No solids N/AEtOH/water 50/50 and acid at 70° C. (clear/colorless). Cool to RT,stirring, 5 days. Mixed suspension of free base in Yellow solution, N/Aacetone/MTBE 70/30 (sticky solids). Stirring, no RT. Added MTBE. solidsMixed hot suspension of free base in Unknown DisorderedTHF/heptane/(excess of heptane) with acid morphology, at 65-70° C.(sticky solids beneath clear some solution). Decanted, added IPA,sonication birefringence/ (cloudy). Stirring, RT, overnight. extinctionMixed hot suspension of free base in Unknown Same as TFE/ACN 30/70 withacid at 70-75° C. (clear, morphology, above, gummy solid). SC to RT withstirring, 1 day. solvent present, slightly birefringence/ improvedextinction crystallinity Mixed hot suspension of free base in UnknownX-ray EtOH/water 60/40 with aq acid at 70° C., by morphology, amorphousdropwise addition (clear). SC to RT, 1 day. opaque, solvent present,some birefringence/ extinction Mixed hot solution (yellow) of free basein No solids N/A EtOH/ACN 30/70 with acid at 70-75° C. (clear,colorless). SC to RT, 1 day, partial evaporation. Sulfuric acid Mixedhot suspension of free base in IPA Unknown X-ray (2:1) with acid at 60°C. (some viscous). Cool to morphology, amorphous + RT, birefringence/free base slurry. extinction inside isotropic particles L-Tartaric EtOH,slurry, RT. Unknown Free base acid morphology, (1:1) birefringence/extinction Mixed hot slurry of free base in ACN with Unknown X-ray acidat 60-70° C. (white and yellow solids). morphology, amorphous +Stirring, SC to RT. Added EtOH (80/20 some free base ACN/EtOH), slurry,RT, overnight. birefringence/ extinction inside isotropic particlesMixed hot suspension of free base in IPA Unknown X-ray with acid at 60°C. (some viscous). Cool to morphology, amorphous + RT, slurry.birefringence/ free base extinction p-TSA Mixed hot EtOH solutions ofacid and free Oily viscous N/A (monohydrate) base at 67° C. Stirring,67° C., 30 min then residue (1:1) RT, 2 hours (less yellow and clearer),sonicated. Added ACN, EtOH/ACN, 40/60. Cooled to −15° C. to −25° C.,overnight. Mixed hot EtOH solutions of acid and free No solids N/A baseat 67° C. Stirring, 67° C., 30 min then RT, 2 hours (less yellow andclearer), sonicated. Added ACN, EtOH/ACN, 40/60. Cooled to −15° C. to−25° C., overnight. Re- dissolved at 60° C., evaporated to about ½volume, SC. Sonicated, RT. Mixed hot solution of free base (yellow) inUnknown X-ray IPA with acid at 65° C. (clear/colorless). morphology,amorphous Stirring, 75° C. then RT, overnight (gummy birefringence/solids). extinction

TABLE 9 Abreviations used. Abbreviations/ Category Acronyms FullName/Description Analytical DSC Differential Scanning CalorimetryTechniques NMR Nuclear Magnetic Resonance TGA Thermogravimetric AnalysisXRPD X-ray Powder Diffraction Crystallization FE Fast evaporationTechniques SC Slow cooling SE Slow evaporation RE Rotary evaporation VDVapor Diffusion Miscellaneous Aq. Aqueous LIMS Laboratory InformationManagement System Min Minutes RT Room (ambient) temperature VF Vacuumfiltration Salt Formers BSA Benzenesulfonic acid EDSA1,2-Ethanedisulfonic acid ESA Ethanesulfonic acid MSA Methanesulfonicacid p-TSA p-Toluenesulfonic acid Solvent ACN Acetonitrile CHCl₃Chloroform EtOAc Ethyl acetate EtOH Ethanol HFIPA HexafluoroisopropanolIPA Isopropanol MCH Methylcyclohexane MeOH Methanol MEK Methyl ethylketone MIBK Methylisobutyl ketone MTBE Methyl-tert-butyl ether NMPN-Methyl-2-pirrolidone PG Propylene glycol TFE Trifluoroethanol THFTetrahydrofuran

Screening with nineteen pharmaceutically accepted salt formersidentified three materials exhibiting unique XRPD patterns. The uniquematerials were produced with oxalic acid, sulfuric acid and malonicacid. Salt screen attempts with α keto-glutaric acid, phosphoric acid,L-tartaric acid and derivatives of sulfonic acid either led to X-rayamorphous materials, non-handleable viscous substance, or produced nosolids. The remainder of acids evaluated in this screen resulted in4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide free base or its physicalmixture with free salt formers. Repeated attempts to produce handleablecrystalline materials with sulfonic acids were not successful.

The potential oxalate was produced by ambient temperature slurry inethanol/methyl-tert-butyl ether (55/45). The sample exhibits a uniqueXRPD pattern consistent with a slightly disordered crystalline material.This potential oxalate was not further investigated.

The potential sulfate was produced using a variety of crystallizationtechniques and solvent systems, either via a salt formation reaction orthrough re-crystallization of the corresponding potential salt. Samplesproduced in this study are overall of poor crystallinity, therefore, thematerial was not further investigated.

Example 5 Preparation of4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide Malonate Form A

4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide free base was reacted at 100 mgscale in an ethanol slurry with malonic acid (1:1 molar ratio). After 1day at room temperature the product was purified by vacuum filtrationand evaporation of the filtrate. The resulting4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate Form A formedcrystalline needles.

In a scale-up experiment4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide free base (1 g) was suspendedin ethanol at ambient temperature with stirring and about 1.1equivalents of malonic acid (268 mg) were added. The mixture was thenstirred at ambient temperature for approximately four days, after whichsolids were collected by vacuum filtration. Out of solids isolatedapproximately 500 mg were vacuum-dried at 45° C. overnight and analyzedby XRPD. The remaining solids were analyzed without drying. Both sampleswere consistent with4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate Form A.

A sample of4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate Form A was analyzed byXRPD (FIG. 7). Peaks shown in FIG. 7 are listed in Table 10, prominentpeaks are listed in Table 11.

TABLE 10 XRPD peaks observed for4-(5-Chloro-2-isopropylaminopyridin-4-yl)- 1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl] amide malonate Form A. 2θ (°) dspace (Å) Intensity (%)  3.0 ± 0.2 29.534 ± 1.976  100  5.2 ± 0.2 16.989± 0.653  37  8.0 ± 0.2 11.108 ± 0.279  7 10.9 ± 0.2 8.144 ± 0.150 6 12.1± 0.2 7.339 ± 0.121 1 13.1 ± 0.2 6.732 ± 0.102 3 13.8 ± 0.2 6.404 ±0.092 1 15.7 ± 0.2 5.646 ± 0.072 10 18.4 ± 0.2 4.822 ± 0.052 5 19.8 ±0.2 4.480 ± 0.045 4 20.5 ± 0.2 4.332 ± 0.042 1 21.0 ± 0.2 4.231 ± 0.0401 21.2 ± 0.2 4.191 ± 0.039 2 21.8 ± 0.2 4.067 ± 0.037 1 22.4 ± 0.2 3.959± 0.035 4 23.1 ± 0.2 3.855 ± 0.033 6 23.7 ± 0.2 3.759 ± 0.031 2 24.0 ±0.2 3.698 ± 0.030 3 24.8 ± 0.2 3.586 ± 0.028 3 25.4 ± 0.2 3.508 ± 0.0275 25.9 ± 0.2 3.435 ± 0.026 3 26.3 ± 0.2 3.384 ± 0.025 1 26.5 ± 0.2 3.364± 0.025 1 26.8 ± 0.2 3.320 ± 0.024 2 27.0 ± 0.2 3.302 ± 0.024 3 27.3 ±0.2 3.260 ± 0.023 3 28.0 ± 0.2 3.183 ± 0.022 3 29.0 ± 0.2 3.076 ± 0.0213 29.5 ± 0.2 3.027 ± 0.020 3 30.3 ± 0.2 2.949 ± 0.019 1 30.9 ± 0.2 2.890± 0.018 1

TABLE 11 Prominent XRPD peaks for4-(5-Chloro-2-isopropylaminopyridin-4-yl)- 1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl] amide malonate Form A. 2θ (°) dspace (Å) Intensity (%) 3.0 ± 0.2 29.534 ± 1.976 100 5.2 ± 0.2 16.989 ±0.653  37 15.7 ± 0.2   5.646 ± 0.072  10

FT-IR was performed on a sample of4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate Form A as described inExample 1 (FIG. 8). Observed peaks from FIG. 8 are listed in Table 12.

TABLE 12 Observed FT-IR peaks for4-(5-Chloro-2-isopropylaminopyridin-4-yl)- 1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl] amide malonate Form A. Position(cm⁻¹) Intensity 678 0.0254 691 0.0622 712 0.0612 749 0.104 787 0.0604797 0.0206 821 0.0184 835 0.0316 883 0.0514 917 0.0115 951 0.0188 9630.0245 988 0.0179 1000 0.0214 1033 0.0419 1070 0.0243 1092 0.0202 11040.0146 1112 0.0132 1125 0.0122 1165 0.0393 1198 0.0237 1228 0.0171 12530.0389 1264 0.0229 1275 0.0285 1280 0.0264 1294 0.0287 1315 0.0185 13450.0513 1363 0.0365 1383 0.0308 1393 0.0331 1425 0.0306 1441 0.0367 14750.0511 1504 0.049 1543 0.0438 1573 0.0548 1598 0.0401 1626 0.0796 16370.0585 1670 0.035 1700 0.0421 1922 0.0074 2435 0.0078 2891 0.0103 29720.0133 3072 0.01 3312 0.0089

DSC was performed on a sample of4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate Form A as described inExample 1 (FIG. 9) and showed a prominent endotherm having an onsettemperature of approximately 142.1° C.

Example 6 Aqueous Dissolution of4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide Malonate Form A

Samples of4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide Form C and4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate Form A were eachshaken at ambient temperature in fasting state simulated gastric fluid(FaSSGF) pH 1.6 for 30 minutes. Concentration of4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide was measured at 5, 15 and 30minutes.

After 30 minutes, the samples were removed from FaSSGF, placed infasting state simulated intestinal fluid (FaSSIF) pH 6.5, with shaking,for an additional 5 hours. Concentration of4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide was measured at 10, 30, 60 90,120, 180, 270, and 300 minutes. Results are summarized in Table 13 andshown in FIG. 10A (FaSSGF) and FIG. 10B (FaSSIF).

TABLE 13 Solubility of4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2- carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide Form C and Malonate Form A.Conc of Malonate Form Conc of Form C Media Time (min) A (ug/mL) (ug/mL)pH 1.6 5 380.63 51.29 FaSSGF buffer 15 385.23 101.35 30 368.82 136.97 pH6.5 FaSSIF 10 63.31 83.07 buffer 30 109.88 169.89 60 114.13 141.33 90115.37 378.87 120 110.00 332.68 180 110.49 123.43 270 115.71 128.38 300115.26 154.70

Example 7 Pharmacokinetic Evaluation of4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide Malonate Form A

8 non-naïve male beagles were given4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide Form C and4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate Form A in a two-waycross-over design with at least a 7 day washout period between doses.Each animal received a single dose of drug at 5 mg/kg by oral gavage andplasma analysis of4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide was performed pre-dose(time=0), at 0.5, 1, 2, 4, 6, 8, 12 and 24 hours. Plasma drugconcentrations are shown in FIG. 11A and the average area under thecurve (AUC) is shown in FIG. 11B. Error bars shown in FIG. 11B arecoefficient of variation (CV) of 39% and 50% for4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide Form C and4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate Form A, respectively.

Although illustrative embodiments of the present invention have beendescribed herein, it should be understood that the invention is notlimited to those described, and that various other changes ormodifications may be made by one skilled in the art without departingfrom the scope or spirit of the invention.

CITED REFERENCES

-   1. Kohno M, Pouyssegur J (2006) Targeting the ERK signaling pathway    in cancer therapy. Ann Med 38: 200-211.-   2. Kuby, J., Immunology, 3rd Ed., W.H. Freeman & Co., New York.-   3. Lee D C, Webb M L (2003) Pharmaceutical Analysis. John Wiley &    Sons, Inc., New York: 255-257.-   4. Peterson M L, Hickey M B, Zaworotko M J and Almarsson O (2006)    Expanding the Scope of Crystal Form Evaluation in Pharmaceutical    Science. J Pharm Pharmaceut Sci 9(3):317-326.-   5. Pierce Catalog and Handbook, 1994-1995; Pierce Chemical Co.,    Rockford, Ill.-   6. Remington, The Science and Practice of Pharmacy (21st Edition,    Lippincott Williams and Wilkins, Philadelphia, Pa.-   7. The United States Pharmacopeia—National Formulary, The United    States Pharmacopeial Convention, Rockville, Md.    All documents cited in this application are hereby incorporated by    reference as if recited in full herein.

What is claimed is:
 1. A malonate salt of a compound of formula (I):

which is a crystalline salt having an X-ray powder diffraction(XRPD)pattern comprising characteristic peaks at about 3.0 and 5.2° 2θ.
 2. Themalonate salt according to claim 1 having an X-ray powder diffraction(XRPD) pattern comprising characteristic peaks selected from the groupconsisting of about 3.0, 5.2, 8.0, and 10.9° 2θ.
 3. The malonate saltaccording to claim 1 having an X-ray powder diffraction (XRPD) patterncomprising XRPD 2θ-reflections)(°) at about 3.0. 5.2, 8.0, 10.9, 15.7,18.4, 23.1, and 25.4.
 4. A malonate salt of a compound of formula (I):

having an XRPD pattern substantially as shown in FIG.
 7. 5. Form Acrystalline4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate according to claim 1having an infrared spectroscopy (IR) spectrum comprising one or morepeaks at about 1573, 1504, 1475, 1253, 1033, and 883 cm⁻¹.
 6. Form Acrystalline4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate according to claim 1having an IR spectrum substantially as shown in FIG.
 8. 7. Form Acrystalline 4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid [1-(3-chlorophenyl)-2-hydroxyethyl]amidemalonate according to claim 1 having (i) an XRPD pattern comprising oneor more peaks at about 3.0. 5.2, 8.0, and 10.9° 2θ; and (ii) an IRspectrum comprising one or more peaks at about 1573, 1504, 1475, 1253,1033, and 883 cm⁻¹.
 8. Form A crystalline4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate according to claim 1having a DSC thermogram with an endotherm having an onset temperature ofapproximately 142.1° C.
 9. Form A crystalline4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyI)-2-hydroxyethyl]amide malonate according to claim 1having a DSC thermogram substantially as shown in FIG.
 9. 10. Apharmaceutical composition comprising a crystalline compound accordingto any one of claims 1 or 4-9 and a pharmaceutically acceptable carrier.11. A method of treating a cancer in a subject in need thereofcomprising administering to the subject an effective amount of acrystalline compound according to any one of claims 1 or 4-9, whereinthe cancer is an ERK-dependent cancer associated with an extracellularsignal-related kinase (ERK) pathway.
 12. The method according to claim11, wherein the subject is a mammal.
 13. The method according to claim12, wherein the mammal is selected from the group consisting of humans,primates, farm animals, and domestic animals.
 14. The method accordingto claim 12, wherein the mammal is a human.
 15. The method according toclaim 11 further comprising administering to the subject at least oneadditional anti-cancer agent.
 16. A method of treating a cancer in asubject in need thereof comprising administering to the subject aneffective amount of a pharmaceutical composition according to claim 10,wherein the cancer is an ERK-dependent cancer associated with anextracellular signal-related kinase (ERK) pathway.
 17. The methodaccording to claim 16, wherein the subject is a mammal.
 18. The methodaccording to claim 17, wherein the mammal is selected from the groupconsisting of humans, primates, farm animals, and domestic animals. 19.The method according to claim 17, wherein the mammal is a human.
 20. Themethod according to claim 16 further comprising administering to thesubject at least one additional anti-cancer agent.
 21. A method ofmaking Form A crystalline4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate comprising reactingmalonic acid and4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide under conditions suitable toform Form A crystalline4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate.
 22. The methodaccording to claim 21, wherein the malonic acid and4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide are reacted in an ethanolslurry.